Functional connectivity of coral reef fishes in a tropical seascape assessed by compound-specific stable isotope analyses

Submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy at the Massachusetts Institute of Technology and the Woods Hole Oceanographic Institution February 2011The ecological integrity of tropical habitats, including mangroves, seagrass beds and coral reefs, is coming under increasing pressure from human activities. Many coral reef fish species are thought to use mangroves and seagrass beds as juvenile nurseries before migrating to coral reefs as adults. Identifying essential habitats and preserving functional linkages among these habitats is likely necessary to promote ecosystem health and sustainable fisheries on coral reefs. This necessitates quantitative assessment of functional connectivity among essential habitats at the seascape level. This thesis presents the development and first application of a method for tracking fish migration using amino acid (AA) δ13C analysis in otoliths. In a controlled feeding experiment with fish reared on isotopically distinct diets, we showed that essential AAs exhibited minimal trophic fractionation between consumer and diet, providing a δ13C record of the baseline isoscape. We explored the potential for geochemical signatures in otoliths of snapper to act as natural tags of residency in seagrass beds, mangroves and coral reefs in the Red Sea, Caribbean Sea and Eastern Pacific Ocean. The δ13C values of otolith essential AAs varied as a function of habitat type and provided a better tracer of residence in juvenile nursery habitats than conventional bulk stable isotope analyses (SIA). Using our otolith AA SIA approach, we quantified the relative contribution of coastal wetlands and reef habitats to Lutjanus ehrenbergii populations on coastal, shelf and oceanic coral reefs in the Red Sea. L. ehrenbergii made significant ontogenetic migrations, traveling more than 30 km from juvenile nurseries to coral reefs and across deep open water. Coastal wetlands were important nurseries for L. ehrenbergii; however, there was significant plasticity in L. ehrenbergii juvenile habitat requirements. Seascape configuration played an important role in determining the functional connectivity of L. ehrenbergii populations in the Red Sea. The compound-specific SIA approach presented in this thesis will be particularly valuable for tracking the movement of species and life-stages not amenable to conventional tagging techniques. This thesis provides quantitative scientific support for establishing realistic population connectivity models that can be used to design effective marine reserve networks.I have been supported by an Ocean Life Institute Fellowship, a National Science Foundation Graduate Research Fellowship, and the WHOI Academic Programs Office. The research presented in this thesis was supported by an Ocean Life Institute student research grant to K. McMahon, an International Society for Reef Studies-Ocean Conservancy Coral Reef Research Fellowship to K. McMahon, and King Abdullah University of Science and Technology (KAUST) Award Nos. USA 00002 and KSA 00011 to S. Thorrold. Additional support came from the Woods Hole Oceanographic Institution, the Large Pelagics Research Center at the University of New Hampshire, the Carnegie Institution of Washington and the W.M. Keck Foundation

A review of ecogeochemistry approaches to estimating movements of marine animals

Author Posting. © Association for the Sciences of Limnology and Oceanography, 2013. This article is posted here by permission of Association for the Sciences of Limnology and Oceanography for personal use, not for redistribution. The definitive version was published in Limnology and Oceanography 58 (2013): 697-714, doi:10.4319/lo.2013.58.2.0697.Ecogeochemistry—the application of geochemical techniques to fundamental questions in population and community ecology—has been used in animal migration studies in terrestrial environments for several decades; however, the approach has received far less attention in marine systems. This review includes comprehensive meta-analyses of organic zooplankton δ13C and δ15N values at the base of the food web, dissolved inorganic carbon δ13C values, and seawater δ18O values to create, for the first time, robust isoscapes for the Atlantic Ocean. These isoscapes present far greater geographic variability in multiple geochemical tracers than was previously thought, thus forming the foundation for reconstructions of habitat use and migration patterns of marine organisms. We review several additional tracers, including trace-element-to-calcium ratios and heavy element stable isotopes, to examine anadromous migrations. We highlight the value of the ecogeochemistry approach by examining case studies on three components of connectivity: dispersal and natal homing, functional connectivity, and migratory connectivity. We also discuss recent advances in compound-specific stable carbon and nitrogen isotope analyses for tracking animal movement. A better understanding of isotopic routing and fractionation factors, particularly of individual compound classes, is necessary to realize the full potential of ecogeochemistry.We were supported by funding from the National Science Foundation (Division of Ocean Sciences–0825148 to S.R.T.), Award USA 00002 and KSA 00011 from the King Abdullah University of Science and Technology (to S.R.T.), and a National Science Foundation Graduate Research Fellowship (to L.H.)

Tracing carbon flow through coral reef food webs using a compound-specific stable isotope approach

Author Posting. © The Author(s), 2015. This is the author's version of the work. It is posted here by permission of Springer for personal use, not for redistribution. The definitive version was published in Oecologia 180 (2016): 809-821, doi:10.1007/s00442-015-3475-3.Coral reefs support spectacularly productive and diverse communities in tropical and sub26 tropical waters throughout the world’s oceans. Debate continues, however, on the degree to which reef biomass is supported by new water column production, benthic primary production, and recycled detrital carbon. We coupled compound-specific δ13C analyses with Bayesian mixing models to quantify carbon flow from primary producers to coral reef fishes across multiple feeding guilds and trophic positions in the Red Sea. Analyses of reef fishes with putative diets composed primarily of zooplankton (Amblyglyphidodon indicus), benthic macroalgae (Stegastes nigricans), reef-associated detritus (Ctenochaetus striatus), and coral tissue (Chaetodon trifascialis) confirmed that δ13C values of essential amino acids from all baseline carbon sources were both isotopically diagnostic and accurately recorded in consumer tissues. While all four source end-members contributed to the production of coral reef fishes in our study, a single source end-member often dominated dietary carbon assimilation of a given species, even for highly mobile, generalist top predators. Microbially-reworked detritus was an important secondary carbon source for most species. Seascape configuration played an important role in structuring resource utilization patterns. For instance, L. ehrenbergii, showed a significant shift from a benthic macroalgal food web on shelf reefs (71 ± 13% of dietary carbon) to a phytoplankton-based food web (72 ± 11%) on oceanic reefs. Our work provides insights into the roles that diverse carbon sources play in the structure and function of coral reef ecosystems and illustrates a powerful fingerprinting method to develop and test nutritional frameworks for understanding resource utilization.This research was based on work supported by Awards USA 00002 and KSA 00011 from King Abdullah University of Science and Technology (KAUST); additional funding was provided by the Woods Hole Oceanographic Institution (WHOI), a KAUST-WHOI award (SPCF-7000000104), and KAUST baseline research funds.2016-11-2

Meta-analysis of primary producer amino acid δ\u3csup\u3e15\u3c/sup\u3eN values and their influence on trophic position estimation

Compound-specific stable isotope analysis of individual amino acids (CSIA-AA) has emerged as a transformative approach to estimate consumer trophic positions (TPCSIA) that are internally indexed to primary producer nitrogen isotope baselines. Central to accurate TPCSIA estimation is an understanding of beta (β) values—the differences between trophic and source AA δ15N values in the primary producers at the base of a consumers’ food web. Growing evidence suggests higher taxonomic and tissue-specific β value variability than typically appreciated. This meta-analysis fulfills a pressing need to comprehensively evaluate relevant sources of β value variability and its contribution to TPCSIA uncertainty. We first synthesized all published primary producer AA δ15N data to investigate ecologically relevant sources of variability (e.g., taxonomy, tissue type, habitat type, mode of photosynthesis). We then reviewed the biogeochemical mechanisms underpinning AA δ15N and β value variability. Lastly, we evaluated the sensitivity of TPCSIA estimates to uncertainty in mean βGlx-Phe values and Glx-Phe trophic discrimination factors (TDFGlx-Phe). We show that variation in βGlx-Phe values is two times greater than previously considered, with degree of vascularization, not habitat type (terrestrial vs. aquatic), providing the greatest source of variability (vascular autotroph = –6.6 ± 3.4‰; non-vascular autotroph = +3.3 ± 1.8‰). Within vascular plants, tissue type secondarily contributed to βGlx-Phe value variability, but we found no clear distinction among C3, C4, and CAM plant βGlx-Phe values. Notably, we found that vascular plant βGlx-Lysvalues (+2.5 ± 1.6‰) are considerably less variable than βGlx-Phe values, making Lys a useful AA tracer of primary production sources in terrestrial systems. Our multi-trophic level sensitivity analyses demonstrate that TPCSIA estimates are highly sensitive to changes in both βGlx-Phe and TDFGlx-Phe values but that the relative influence of β values dissipates at higher trophic levels. Our results highlight that primary producer β values are integral to accurate trophic position estimation. We outline four key recommendations for identifying, constraining, and accounting for β value variability to improve TPCSIA estimation accuracy and precision moving forward. We must ultimately expand libraries of primary producer AA δ15N values to better understand mechanistic drivers of β value variation

Major shifts in nutrient and phytoplankton dynamics in the North Pacific Subtropical Gyre over the last 5000 years revealed by high-resolution proteinaceous deep-sea coral δ\u3csup\u3e15\u3c/sup\u3eN and δ\u3csup\u3e13\u3c/sup\u3eC records

The North Pacific Subtropical Gyre (NPSG) is the largest continuous ecosystem on Earth and is a critical component of global oceanic biogeochemical cycling and carbon sequestration. We report here multi-millennial-scale, sub-decadal-resolution records of bulk stable nitrogen (δ15N) and carbon (δ13C) isotope records from proteinaceous deep-sea corals. Data from three Kulamanamana haumeaae specimens from the main Hawaiian Islands extend the coral-based time-series back ∼5000 yrs for the NPSG and bypass constraints of low resolution sediment cores in this oligotrophic ocean region. We interpret these records in terms of shifting biogeochemical cycles and plankton community structure, with a main goal of placing the extraordinarily rapid ecosystem biogeochemical changes documented by recent coral records during the Anthropocene in a context of broader Late-Holocene variability. During intervals where new data overlaps with previous records, there is strong correspondence in isotope values, indicating that this older data represents a direct extension of Anthropocene records. These results reveal multiple large isotopic shifts in both δ15N and δ13C values similar to or larger in magnitude to those reported in the last 150 yrs. This shows that large fluctuations in the isotopic composition of export production in this region are not unique to the recent past, but have occurred multiple times through the Mid- to Late-Holocene. However, these earlier isotopic shifts occurred over much longer time intervals (∼millennial vs. decadal timescales). Further, the δ15N data confirm that the extremely low present day δ15N values recorded by deep sea corals (∼8‰) are unprecedented for the NPSG, at least within the past five millennia. Together these records reveal centennial to millennial-scale oscillations in NPSG biogeochemical cycles. Further, these data also suggest a number of independent biogeochemical regimes during which δ15N and δ13C trends were synchronous (similar to recent coral records) or distinctly decoupled. We propose that phytoplankton species composition and nutrient source changes are the dominant mechanisms controlling the coupling and de-coupling of δ15N and δ13C values, likely primarily influenced by changing oceanographic conditions (e.g., stratification vs. entrainment). The decoupling observed in the past further suggests that oceanographic forcing and ecosystem responses controlling δ15N and δ13C values of export production have been substantially different earlier in the Holocene compared to mechanisms controlling the present day system

Carbon and nitrogen isotope fractionation of amino acids in an avian marine predator, the gentoo penguin (Pygoscelis papua)

© The Author(s), 2015. This article is distributed under the terms of the Creative Commons Attribution License. The definitive version was published in Ecology and Evolution 5 (2015): 1278–1290, doi:10.1002/ece3.1437.Compound-specific stable isotope analysis (CSIA) of amino acids (AA) has rapidly become a powerful tool in studies of food web architecture, resource use, and biogeochemical cycling. However, applications to avian ecology have been limited because no controlled studies have examined the patterns in AA isotope fractionation in birds. We conducted a controlled CSIA feeding experiment on an avian species, the gentoo penguin (Pygoscelis papua), to examine patterns in individual AA carbon and nitrogen stable isotope fractionation between diet (D) and consumer (C) (Δ13CC-D and Δ15NC-D, respectively). We found that essential AA δ13C values and source AA δ15N values in feathers showed minimal trophic fractionation between diet and consumer, providing independent but complimentary archival proxies for primary producers and nitrogen sources respectively, at the base of food webs supporting penguins. Variations in nonessential AA Δ13CC-D values reflected differences in macromolecule sources used for biosynthesis (e.g., protein vs. lipids) and provided a metric to assess resource utilization. The avian-specific nitrogen trophic discrimination factor (TDFGlu-Phe = 3.5 ± 0.4‰) that we calculated from the difference in trophic fractionation (Δ15NC-D) of glutamic acid and phenylalanine was significantly lower than the conventional literature value of 7.6‰. Trophic positions of five species of wild penguins calculated using a multi-TDFGlu-Phe equation with the avian-specific TDFGlu-Phe value from our experiment provided estimates that were more ecologically realistic than estimates using a single TDFGlu-Phe of 7.6‰ from the previous literature. Our results provide a quantitative, mechanistic framework for the use of CSIA in nonlethal, archival feathers to study the movement and foraging ecology of avian consumers.This research was funded by National Science Foundation Office of Polar Programs [grants ANT-0125098, ANT-0739575] and the 2013 Antarctic Science Bursaries

Linking habitat mosaics and connectivity in a coral reef seascape

Author Posting. © The Author(s), 2012. This is the author's version of the work. It is posted here by permission of National Academy of Sciences for personal use, not for redistribution. The definitive version was published in Proceedings of the National Academy of Sciences of the United States of America 109 (2012): 15372-15376, doi:10.1073/pnas.1206378109.Tropical marine ecosystems are under mounting anthropogenic pressure from overfishing and habitat destruction, leading to declines in their structure and function on a global scale. While maintaining connectivity among habitats within a seascape is necessary for preserving population resistance and resilience, quantifying movements of individuals within seascapes remains challenging. Traditional methods of identifying and valuing potential coral reef fish nursery habitats are indirect, often relying on visual surveys of abundance and correlations of size and biomass among habitats. We used compound-specific stable isotope analyses to determine movement patterns of commercially important fish populations within a coral reef seascape. This approach allowed us to quantify the relative contributions of individuals from inshore nurseries to reef populations and identify migration corridors among important habitats. Our results provided direct measurements of remarkable migrations by juvenile snapper of over 30 km between nurseries and reefs. We also found significant plasticity in juvenile nursery residency. While a majority of individuals on coastal reefs had used seagrass nurseries as juveniles, many adults on oceanic reefs had settled directly into reef habitats. Moreover, seascape configuration played a critical but heretofore unrecognized role in determining connectivity among habitats. Finally, our approach provides key quantitative data necessary to estimate the value of distinctive habitats to ecosystem services provided by seascapes.This research was based on work supported by Award Nos. USA 00002 and KSA 00011 made by King Abdullah University of Science and Technology (KAUST). Additional funding was provided by Woods Hole Oceanographic Institution and an International Society for Reef Studies-Ocean Conservancy Coral Reef Fellowship. K. McMahon received support from the National Science Foundation Graduate Research Fellowship Program

Calibrating amino acid δ\u3csup\u3e13\u3c/sup\u3eC and δ\u3csup\u3e15\u3c/sup\u3eN offsets between polyp and protein skeleton to develop proteinaceous deep-sea corals as paleoceanographic archives.

Compound-specific stable isotopes of amino acids (CSI-AA) from proteinaceous deep-sea coral skeletons have the potential to improve paleoreconstructions of plankton community composition, and our understanding of the trophic dynamics and biogeochemical cycling of sinking organic matter in the Ocean. However, the assumption that the molecular isotopic values preserved in protein skeletal material reflect those of the living coral polyps has never been directly investigated in proteinaceous deep-sea corals. We examined CSI-AA from three genera of proteinaceous deep-sea corals from three oceanographically distinct regions of the North Pacific: Primnoa from the Gulf of Alaska, Isidella from the Central California Margin, and Kulamanamana from the North Pacific Subtropical Gyre. We found minimal offsets in the δ13C values of both essential and non-essential AAs, and in the δ15N values of source AAs, between paired samples of polyp tissue and protein skeleton. Using an essential AA δ13C fingerprinting approach, we show that estimates of the relative contribution of eukaryotic microalgae and prokaryotic cyanobacteria to the sinking organic matter supporting deep-sea corals are the same when calculated from polyp tissue or recently deposited skeletal tissue. The δ15N values of trophic AAs in skeletal tissue, on the other hand, were consistently 3–4‰ lower than polyp tissue for all three genera. We hypothesize that this offset reflects a partitioning of nitrogen flux through isotopic branch points in the synthesis of polyp (fast turnover tissue) and skeleton (slow, unidirectional incorporation). This offset indicates an underestimation, albeit correctable, of approximately half a trophic position from gorgonin protein-based deep-sea coral skeleton. Together, our observations open the door for applying many of the rapidly evolving CSI-AA based tools developed for metabolically active tissues in modern systems to archival coral tissues in a paleoceanographic context

Impact of skeletal heterogeneity and treatment method on interpretation of environmental variability from the proteinaceous skeletons of deep-sea gorgonian octocorals

The stable isotope geochemistry of gorgonian octocoral skeletons facilitates detailed time series reconstructions of nutrient biogeochemistry. However, comparisons among reconstructions from different locations require realistic estimates of the uncertainty surrounding each measured geochemical value. Here, we determine quantitative uncertainties related to 1) standard skeletal pretreatment in preparation for stable isotopic analysis and 2) biological variability associated with a heterogeneous isotopic composition of the gorgonin skeleton. We found that the 5% HCl pretreatment required for the δ13C measurements does not significantly impact the δ15N values of the skeleton nor the reproducibility of the δ15N measurements. In contrast, while 5% HCl pretreatment significantly altered bulk δ13C values via removal of CaCO3, it did not change amino acid δ13C values in the organic skeleton. We found that the variance of repeat measurements of skeleton samples formed contemporaneously and homogenized skeleton for both δ13C and δ15N exceeded that of instrumental uncertainty of an acetanilide standard. This indicates that instrumental uncertainty underestimates the true precision of an isotopic measurement of the organic skeleton. Furthermore, measurements of contemporaneous skeleton around the circumference of an octocoral colony yielded variability exceeding that of homogenized skeleton. Based on these results, we find that 1) both δ13C and δ15N values can be measured simultaneously in pretreated skeleton, 2) growth bands should be homogenized prior to analysis, and 3) reported error should include uncertainty due to biological effects determined from repeat analysis of homogenized skeleton and not just instrument error to reduce false significant differences. Our results present an important protocol for processing proteinaceous octocoral skeletons and propagating uncertainty to more accurately reconstruct nutrient dynamics from proteinaceous deep-sea octocoral skeletons

Amino acid isotope discrimination factors for a carnivore: physiological insights from leopard sharks and their diet

Stable isotopes are important ecological tools, because the carbon and nitrogen isotopic composition of consumer tissue reflects the diet. Measurements of isotopes of individual amino acids can disentangle the effects of consumer physiology from spatiotemporal variation in dietary isotopic values. However, this approach requires knowledge of assimilation patterns of dietary amino acids. We reared leopard sharks (Triakis semifasciata) on diets of squid (Loligo opalescens; 1250 days; control sharks) or squid then tilapia (Oreochromis sp.; switched at 565 days; experimental sharks) to evaluate consumer-diet discrimination factors for amino acids in muscle tissue. We found that control sharks exhibited lower nitrogen isotope discrimination factors (∆15N) than most previous consumer studies, potentially because of urea recycling. Control sharks also had large carbon isotope discrimination factors (∆13C) for three essential amino acids, suggesting microbial contributions or fractionation upon assimilation. Compared to controls, experimental sharks exhibited higher ∆13C values for four amino acids and ∆15N values for seven amino acids, corresponding with differences between diets in δ13C and δ15N values. This suggests that not all amino acids in experimental sharks had reached steady state, contrary to the conclusion of a bulk isotope study of these sharks. Our results imply that (1) the magnitude of a shift in dietary δ13C and δ15N values temporarily influences the appearance of discrimination factors; (2) slow turnover of amino acid isotopes in elasmobranch muscle precludes inferences about seasonal dietary changes; (3) elasmobranch discrimination factors for amino acids may be affected by urea recycling and microbial contributions of amino acids