AbstractFindings from eDNA metabarcoding are strongly influenced by experimental approach, yet the effect of pre‐PCR sample processing on taxon detection and estimates of biodiversity across different water types is still poorly resolved. To fill this data gap, we investigated the impact of sampling effort, extraction method, and filter pore size on DNA yield, PCR inhibition, and 16S rDNA metabarcoding results for fishes in water samples collected from inshore turbid‐ and offshore clear‐water environments. The turbid‐water samples had high concentrations of suspended organic and/or inorganic material and yielded ~3.2× more DNA and exhibited high levels of PCR inhibition compared with the low‐turbidity, clear‐water samples. Importantly, there were no striking differences in the results of our metabarcoding experiments based on extraction method or filter pore size. While a small number of unique species of relatively low read count were detected in all turbid‐water treatments, most species were consistently detected across samples. Results for the clear‐water samples were strikingly different, with low DNA yield, high levels of variation across replicates, and a high number of non‐overlapping species across treatments. These findings indicate a patchy distribution of eDNA in offshore environments, which means higher volumes of water (≥ 2 L per replicate) must be filtered in habitats where target DNA is likely to be sparse. In semi‐closed systems such as estuaries, higher concentrations of target DNA are expected, and we found that either a 1.0 or 3.0 µm filter pore size was sufficient to capture standing diversity, while decreasing the risk of clogging. For economical DNA extraction and inhibitor removal, we recommend a combination of Omega Bio‐tek E.Z.N.A Tissue DNA kit followed by a PCR inhibitor removal step using the Zymo Kit. Finally, we emphasize that pilot studies should be undertaken whenever sampling in a new environment to identify which protocol is most appropriate.

Environmental DNA (eDNA) is increasingly being used to document species distributions and habitat use in marine systems, with much of the recent effort focused on leveraging advances in next-generation DNA sequencing to assess and track biodiversity across taxonomic groups. Environmental DNA offers a number of important advantages over traditional survey techniques, including non-invasive sampling, sampling where traditional approaches are impractical or inefficient (e.g. deep oceans), reduced cost, and increased detection sensitivity. However, eDNA applications are currently limited because of an insufficient understanding of the influence of sample source, analytical approach, and marker type on eDNA detections. Because approaches vary considerably among eDNA studies, we present a summary of the current state of the field and emerging best practices. The impact of observed variation in rates of eDNA production, persistence, and transport are also discussed and future research needs are highlighted with the goal of expanding eDNA applications, including the development of statistical models to improve the predictability of eDNA detection and quantification.

Aim To test hypothesized biogeographic partitions of the tropical Indo-Pacific Ocean with phylogeographic data from 56 taxa, and to evaluate the strength and nature of barriers emerging from this test. Location The Indo-Pacific Ocean. Time period Pliocene through the Holocene. Major taxa studied Fifty-six marine species. Methods We tested eight biogeographic hypotheses for partitioning of the Indo-Pacific using a novel modification to analysis of molecular variance. Putative barriers to gene flow emerging from this analysis were evaluated for pairwise phi(ST), and these phi(ST) distributions were compared to distributions from randomized datasets and simple coalescent simulations of vicariance arising from the Last Glacial Maximum. We then weighed the relative contribution of distance versus environmental or geographic barriers to pairwise phi(ST) with a distance-based redundancy analysis (dbRDA). Results We observed a diversity of outcomes, although the majority of species fit a few broad biogeographic regions. Repeated coalescent simulation of a simple vicariance model yielded a wide distribution of pairwise phi(ST) that was very similar to empirical distributions observed across five putative barriers to gene flow. Three of these barriers had median phi(ST) that were significantly larger than random expectation. Only 21 of 52 species analysed with dbRDA rejected the null model. Among these, 15 had overwater distance as a significant predictor of pairwise phi(ST), while 11 were significant for geographic or environmental barriers other than distance. Main conclusions Although there is support for three previously described barriers, phylogeographic discordance in the Indo-Pacific Ocean indicates incongruity between processes shaping the distributions of diversity at the species and population levels. Among the many possible causes of this incongruity, genetic drift provides the most compelling explanation: given massive effective population sizes of Indo-Pacific species, even hard vicariance for tens of thousands of years can yield phi(ST )values that range from 0 to nearly 0.5.

Student participation in authentic research, as citizen scientists, can improve classroom engagement, achievement of learning objectives, and perceptions of science. We present DNA barcoding of invasive lionfish (Pterois volitans) prey as an example student citizen-science project, though the protocols, objectives, and outcomes can be generalized to any piscivorous fish. The objective of this five-lab conservation genetics unit is to enhance student understanding of fundamental molecular and ecological concepts through applied use of DNA sequencing technologies. Student assessments were equivocal, indicating modest gains in conceptual understanding and maintenance of an overall high perception of science. More notably, student findings have contributed to an improved understanding of the impacts of invasive lionfish, including providing the first evidence that lionfish prey on economically important red snapper (Lutjanus campechanus).

Bluespine unicornfish (Naso unicornis) from Hawaii were aged to >50 years using cross-sectioned sagittal otoliths. Fish length was a poor indicator of age because of rapid and variable early growth, exemplified by fish aged to be 4 years near maximum length. Growth was deterministic with adult ages decoupled from body length. Otolith mass and thickness were evaluated as proxies for age and both were encouraging; thickness explained more variance but mass was easier to measure. An age estimation protocol was validated through ontogeny using bomb radiocarbon (14C) dating. Use of the postbomb 14C decline period from a regional reference chronology enabled age validation of young fish — a novel approach for the Pacific Ocean. A probabilistic procedure for assigning bomb 14C dates (CALIBomb) was used for the first time to determine fish birth years. The age-reading protocol was generally validated, and it was possible to describe length-at-age despite difficulties in counting otolith annuli beyond 30–40 years. Growth curves differed between the sexes, and a four-parameter generalized von Bertalanffy growth function provided the best fit.

The regal angelfish (Pygoplites diacanthus; family Pomacanthidae) occurs on reefs from the Red Sea to the central Pacific, with an Indian Ocean/Rea Sea color morph distinct from a Pacific Ocean morph. To assess population differentiation and evaluate the possibility of cryptic evolutionary partitions in this monotypic genus, we surveyed mtDNA cytochrome b and two nuclear introns (S7 and RAG2) in 547 individuals from 15 locations. Phylogeographic analyses revealed four mtDNA lineages (d=0.006-0.015) corresponding to the Pacific Ocean, the Red Sea, and two admixed lineages in the Indian Ocean, a pattern consistent with known biogeographic barriers. Christmas Island in the eastern Indian Ocean had both Indian and Pacific lineages. Both S7 and RAG2 showed strong population-level differentiation between the Red Sea, Indian Ocean, and Pacific Ocean (ΦST=0.066-0.512). The only consistent population sub-structure within these three regions was at the Society Islands (French Polynesia), where surrounding oceanographic conditions may reinforce isolation. Coalescence analyses indicate the Pacific (1.7Ma) as the oldest extant lineage followed by the Red Sea lineage (1.4Ma). Results from a median-joining network suggest radiations of two lineages from the Red Sea that currently occupy the Indian Ocean (0.7-0.9Ma). Persistence of a Red Sea lineage through Pleistocene glacial cycles suggests a long-term refuge in this region. The affiliation of Pacific and Red Sea populations, apparent in cytochrome b and S7 (but equivocal in RAG2) raises the hypothesis that the Indian Ocean was recolonized from the Red Sea, possibly more than once. Assessing the genetic architecture of this widespread monotypic genus reveals cryptic evolutionary diversity that merits subspecific recognition. We recommend P.d. diacanthus and P.d. flavescens for the Pacific and Indian Ocean/Red Sea forms.

The few studies that have addressed past effects of climate change on species distributions have mostly focused on plants due to the rarity of historical faunal baselines. However, hyperdiverse groups like Arthropoda are vital to monitor in order to understand climate change impacts on biodiversity. This is the first investigation of ground-dwelling arthropod (GDA) assemblages along the full elevation gradient of a mountain range in the Madrean Sky Island Region, establishing a baseline for monitoring future changes in GDA biodiversity. To determine how GDA assemblages relate to elevation, season, abiotic variables, and corresponding biomes, GDA were collected for two weeks in both spring (May) and summer (September) 2011 in the Santa Catalina Mountains, Arizona, using pitfall traps at 66 sites in six distinct upland (non-riparian/non-wet canyon) biomes. Four arthropod taxa: (1) beetles (Coleoptera), (2) spiders (Araneae), (3) grasshoppers and crickets (Orthoptera), and (4) millipedes and centipedes (Myriapoda) were assessed together and separately to determine if there are similar patterns across taxonomic groups. We collected 335 species of GDA: 192/3793 (species/specimens) Coleoptera, 102/1329 Araneae, 25/523 Orthoptera, and 16/697 Myriapoda. GDA assemblages differed among all biomes and between seasons. Fifty-three percent (178 species) and 76% (254 species) of all GDA species were found in only one biome and during only one season, respectively. While composition of arthropod assemblages is tied to biome and season, individual groups do not show fully concordant patterns. Seventeen percent of the GDA species occurred only in the two highest-elevation biomes (Pine and Mixed Conifer Forests). Because these high elevation biomes are most threatened by climate change and they harbor a large percentage of unique arthropod species (11-25% depending on taxon), significant loss in arthropod diversity is likely in the Santa Catalina Mountains and other isolated mountain ranges in the Southwestern US.

David Bertelsen recently published an essay (Bertelse n 2014) discussing some methodological aspects of plant ecology. The methods he addresse d hav long been debated in the ecological literature and there is no need to comment on them here. However, his essay also critiqued our recent paper on plant distributions and climate change in the Santa Catalina Mountains (Brusca et al. 2014) and, incidentally, also the work of Robert Whittake r and William Niering (Whittaker & Niering 1964; Whittaker 1967). We feel that Mr. Bertelsen’s s ummary of our paper was inaccurate and misleading, and we take this opportunity to correct his misunderstanding of our work.

We herein evaluate several reproductive metrics of Hawaiian Archipelagic populations of the bluespine unicornfish (Naso unicornis), an economically and ecologically important, broadly distributed tropical Pacific reef fish, based on multi-year, fishery-dependent and fishery-independent collections. Sex-specific spawning seasonality was characterized for fish collected mostly from Oahu (Main Hawaiian Islands, MHI) using a gonadosomatic index. Histological slides preparations were used to score gonad developmental phase and to classify individuals of either sex as immature or mature. Sex-specific median body lengths at maturity (L50) were estimated by logistic fits of proportion mature versus length class. Spawning was highly seasonal in Hawaii, with a single brief (May–June) peak spawning period. Proportionate gonad-to-body weight values were relatively low, averaging only about 0.1 % and 0.6 % across all months of year and 0.16 % and 1.03 % during May–June for males and females, respectively. Median lengths at sexual maturity differed between the sexes. L50 values for fish collected throughout all months of year were 30.1 ± 0.5 (standard error) cm Fork Length (FL) for males and 35.5 ± 0.7 cm FL for females. Spawning seasonality and L50 estimates for bluespine unicornfish in Hawaii suggest that the species spawns several months earlier in the calendar year and matures at larger body lengths in Hawaii versus Guam, the Northern Mariana Islands, and Pohnpei in the Federated States of Micronesia. Estimated lengths at sexual maturity are compared to the minimum length (14 inches or 35.6 cm FL) mandated for this species in Hawaii: median size at maturity occurs at a length appreciably less than (males) or approximately equal to (females) minimum legal size. A likely disproportionately large contribution of old females to population replenishment is discussed relative to the minimum size limit.

The Arizona Sky Island Arthropod Project (ASAP) is a new multi-disciplinary research program at the University of Arizona that combines systematics, biogeography, ecology, and population genetics to study origins and patterns of arthropod diversity along elevation gradients and among mountain ranges in the Madrean Sky Island Region. Arthropods represent taxonomically and ecologically diverse organisms that drive key ecosystem processes in this mountain archipelago. Using data from museum specimens and specimens we obtain during long-term collecting and monitoring programs, ASAP will document arthropod species across Arizona's Sky Islands to address a number of fundamental questions about arthropods of this region. Baseline data will be used to determine climatic boundaries for target species, which will then be integrated with climatological models to predict future changes in arthropod communities and distributions in the wake of rapid climate change. ASAP also makes use of the natural laboratory provided by the Sky Islands to investigate ecological and genetic factors that influence diversification and patterns of community assembly. Here, we introduce the project, outline overarching goals, and describe preliminary data from the first year of sampling ground-dwelling beetles and ants in the Santa Catalina Mountains.

Aim The Red Sea is a biodiversity hotspot characterized by a unique marine fauna and high endemism. This sea began forming c.24million years ago with the separation of the African and Arabian plates, and has been characterized by periods of desiccation, hypersalinity and intermittent connection to the Indian Ocean. We aim to evaluate the impact of these events on the genetic architecture of the Red Sea reef fish fauna. Location Red Sea and Western Indian Ocean. Methods We surveyed seven reef fish species from the Red Sea and adjacent Indian Ocean using mitochondrial DNA cytochrome c oxidase subunit I and cytochrome b sequences. To assess genetic variation and evolutionary connectivity within and between these regions, we estimated haplotype diversity (h) and nucleotide diversity (), reconstructed phylogenetic relationships among haplotypes, and estimated gene flow and time of population separation using Bayesian coalescent-based methodology. Results Our analyses revealed a range of scenarios from shallow population structure to diagnostic differences that indicate evolutionary partitions and possible cryptic species. Conventional molecular clocks and coalescence analyses indicated time-frames for divergence between these bodies of water ranging from 830,000years to contemporary exchange or recent range expansion. Colonization routes were bidirectional, with some species moving from the Indian Ocean to the Red Sea compared with expansion out of the Red Sea for other species. Main conclusions We conclude that: (1) at least some Red Sea reef fauna survived multiple salinity crises; (2) endemism is higher in the Red Sea than previously reported; and (3) the Red Sea is an evolutionary incubator, occasionally contributing species to the adjacent Indian Ocean. The latter two conclusions elevated endemism and species export indicate a need for enhanced conservation priorities for the Red Sea.

Models analyzing how Southwestern plant communities will respond to climate change predict that increases in temperature will lead to upward elevational shifts of montane species. We tested this hypothesis by reexamining Robert Whittaker's 1963 plant transect in the Santa Catalina Mountains of southern Arizona, finding that this process is already well underway. Our survey, five decades after Whittaker's, reveals large changes in the elevational ranges of common montane plants, while mean annual rainfall has decreased over the past 20 years, and mean annual temperatures increased 0.25°C/decade from 1949 to 2011 in the Tucson Basin. Although elevational changes in species are individualistic, significant overall upward movement of the lower elevation boundaries, and elevational range contractions, have occurred. This is the first documentation of significant upward shifts of lower elevation range boundaries in Southwestern montane plant species over decadal time, confirming that previous hypotheses are correct in their prediction that mountain communities in the Southwest will be strongly impacted by warming, and that the Southwest is already experiencing a rapid vegetation change.

The Chagos Archipelago was designated a no-take marine protected area (MPA) in 2010; it covers 550 000 km2, with more than 60 000 km2 shallow limestone platform and reefs. This has doubled the global cover of such MPAs.It contains 25-50% of the Indian Ocean reef area remaining in excellent condition, as well as the world's largest contiguous undamaged reef area. It has suffered from warming episodes, but after the most severe mortality event of 1998, coral cover was restored after 10 years.Coral reef fishes are orders of magnitude more abundant than in other Indian Ocean locations, regardless of whether the latter are fished or protected.Coral diseases are extremely low, and no invasive marine species are known.Genetically, Chagos marine species are part of the Western Indian Ocean, and Chagos serves as a 'stepping-stone' in the ocean.The no-take MPA extends to the 200 nm boundary, and. includes 86 unfished seamounts and 243 deep knolls as well as encompassing important pelagic species.On the larger islands, native plants, coconut crabs, bird and turtle colonies were largely destroyed in plantation times, but several smaller islands are in relatively undamaged state.There are now 10 'important bird areas', coconut crab density is high and numbers of green and hawksbill turtles are recovering.Diego Garcia atoll contains a military facility; this atoll contains one Ramsar site and several 'strict nature reserves'. Pollutant monitoring shows it to be the least polluted inhabited atoll in the world. Today, strict environmental regulations are enforced.Shoreline erosion is significant in many places. Its economic cost in the inhabited part of Diego Garcia is very high, but all islands are vulnerable.Chagos is ideally situated for several monitoring programmes, and use is increasingly being made of the archipelago for this purpose.

Determining the geographic scale at which to apply ecosystem-based management (EBM) has proven to be an obstacle for many marine conservation programs. Generalizations based on geographic proximity, taxonomy, or life history characteristics provide little predictive power in determining overall patterns of connectivity, and therefore offer little in terms of delineating boundaries for marine spatial management areas. Here, we provide a case study of 27 taxonomically and ecologically diverse species (including reef fishes, marine mammals, gastropods, echinoderms, cnidarians, crustaceans, and an elasmobranch) that reveal four concordant barriers to dispersal within the Hawaiian Archipelago which are not detected in single-species exemplar studies. We contend that this multispecies approach to determine concordant patterns of connectivity is an objective and logical way in which to define the minimum number of management units and that EBM in the Hawaiian Archipelago requires at least five spatially managed regions.

The depauperate marine ecosystems of the Hawaiian Archipelago share a high proportion of species with the southern and western Pacific, indicating historical and/or ongoing connections across the large oceanic expanse separating Hawaii from its nearest neighbors. The rate and direction of these interactions are, however, unknown. While previous biogeographic studies have consistently described Hawaii as a diversity sink, prevailing currents likely offer opportunities for larval export. To assess interactions between the remote reefs of the Hawaiian Archipelago and the species rich communities of the Central and West Pacific, we surveyed 14 nuclear microsatellite loci (nDNA; n = 857) and a 614 bp segment of mitochondrial cytochrome b (mtDNA; n = 654) in the Yellow Tang (Zebrasoma flavescens). Concordant frequency shifts in both nDNA and mtDNA reveal significant population differentiation among three West Pacific sites and Hawaii (nDNA F' CT = 0.116, mtDNA ϕ CT = 0.098, P < 0.001). SAMOVA analyses of microsatellite data additionally indicate fine scale differentiation within the 2600 km Hawaiian Archipelago (F' SC = 0.026; P < 0.001), with implications for management of this heavily-exploited aquarium fish. Mismatch analyses indicate the oldest contemporary populations are in the Hawaiian Archipelago (circa 318,000 y), with younger populations in the West Pacific (91,000 - 175,000 y). Estimates of Yellow Tang historical demography contradict expectations of Hawaii as a population sink, and instead indicate asymmetrical gene flow, with Hawaii exporting rather than importing Yellow Tang larvae.

Recent reports of localized larval recruitment in predominately small-range fishes are countered by studies that show high genetic connectivity across large oceanic distances. This discrepancy may result from the different timescales over which genetic and demographic processes operate or rather may indicate regular long-distance dispersal in some species. Here, we contribute an analysis of mtDNA cytochrome b diversity in the widely distributed Brown Surgeonfish (Acanthurus nigrofuscus; N = 560), which revealed significant genetic structure only at the extremes of the range (ΦCT = 0.452; P < .001). Collections from Hawaii to the Eastern Indian Ocean comprise one large, undifferentiated population. This pattern of limited genetic subdivision across reefs of the central Indo-Pacific has been observed in a number of large-range reef fishes. Conversely, small-range fishes are often deeply structured over the same area. These findings demonstrate population connectivity differences among species at biogeographic and evolutionary timescales, which likely translates into differences in dispersal ability at ecological and demographic timescales. While interspecific differences in population connectivity complicate the design of management strategies, the integration of multiscale connectivity patterns into marine resource planning will help ensure long-term ecosystem stability by preserving functionally diverse communities.