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Management of protected species is difficult when objectives include the recovery of both predator and its prey. Ideally, identifying trade-offs between competing objectives involves evaluating management alternatives with a quantitative model that integrates information on both species, but data are often limited. We used new predator diet data and simulation modeling to update our understanding of seal predation on juvenile Chinook (Oncorhynchus tshawytscha) and coho salmon (Oncorhynchus kisutch) in the Puget Sound. Under prey size assumptions used by previous studies, estimates suggest predation could be a significant source of mortality for Chinook salmon (mean: 37% of ocean age-0 juveniles), but varied considerably among years (range of median estimates: 22%–49%). However, when we estimated prey size from otoliths recovered from seal scats, the impact (numbers of fish consumed) decreased by 71%. Predation on coho salmon was estimated to be relatively low under both scenarios (6%–8% of ocean age-0 juveniles) with a 21% decrease in consumption using otolith-derived prey size. Our analysis highlights the importance of updating model inputs and re-evaluating assumptions of multi-species models used for ecosystem-based fisheries management.

There is increasing evidence that predation by harbour seals on out-migrating salmon smolts may be responsible for the low return of adult coho and Chinook salmon in the Salish Sea. However, little attention has been given to understanding where and when this predation occurs and the extent to which it might be conducted by few or many seals in the population. We equipped 17 harbour seals with data loggers to track seal movements and used accelerometry to infer prey encounter events (PEEs) following the release of ~384000 coho (May 4, 2015) and ~3 million Chinook (May 14, 2015) smolts into the Big Qualicum River. We found a small proportion (5.7%) of all PEEs occurred in the estuary where salmon smolts entered the ocean—and that only one-quarter of the seals actively fed there. PEE counts increased in the estuary after both species of smolts were released. However, the response of the seals was less synchronous and occurred over a greater range of depths following the release of the smaller-bodied and more abundant Chinook smolts. Harbour seals feeding in the estuary appeared to target coho smolts at the beginning of May but appeared to pursue predators of Chinook smolts in mid-May. PEE counts in the estuary increased as tide height rose and were higher at dusk and night—especially during full moonlight. Such fine-scale behavioural information about harbour seals in relation to pulses of out-migrating smolts can be used to design mitigation strategies to reduce predation pressure by seals on salmon populations.

Environmental DNA (eDNA) detection of aquatic invasive species using quantitative polymerase chain reaction (qPCR) is a powerful tool for resource managers, but qPCR has traditionally been confined to laboratory analysis. Laboratory results often take days or weeks to be produced, limiting options for rapid management response. To circumvent laboratory delay, we combined a purpose‐built eDNA filtration system (Smith‐Root eDNA‐Sampler) with a field DNA extraction and qPCR analysis platform (Biomeme) to form a complete field eDNA sampling and detection process. A controlled laboratory study involving serial dilutions of New Zealand mudsnail (Potamopyrgus antipodarum; Gray, 1843) eDNA was conducted to compare the detection capabilities of the field system with traditional bench qPCR. Additionally, field validation studies were conducted to determine whether field eDNA analysis can be used to map mudsnail eDNA distribution and quantify temporal fluctuations. In the laboratory experiment, both qPCR platforms (Biomeme, bench qPCR) lost the ability to reliably detect mudsnail eDNA at the same dilution level, with starting quantity values as low as 21 DNA copies/reaction. A strong linear relationship was observed between the average quantification cycle values of the two platforms (slope = 1.101, intercept = −1.816, R2 = 0.997, p

With the rise of environmental DNA as a surveillance tool for aquatic species, a need has also arisen for professionally engineered research tools specifically designed for eDNA applications. We created the first portable, purpose-built eDNA sampling system in the form of a backpack smart-pump filtration apparatus and custom made eDNA filter packets for each sample. The eDNA-Sampler (previously ANDe) enables both point-location sampling and mobile sampling over a spatial distance, with the ability to standardize filtration parameters (e.g. flow rate, pressure, water volume, etc.). In this presentation we will describe the evolution of the eDNA sampling backpack and associated components, each designed to help streamline the eDNA sampling process and increase sampling efficiency. We have optimized the platform for mobile sampling by integrating GPS and data logging capabilities, in addition to modifying the chemistry of the eDNA filter packets to minimize the effort required for sample preservation. Results will be presented from a series of pilot studies in which the eDNA-Sampler capabilities were evaluated. Combined, the innovations described herein should help remove barriers to entry for potential eDNA practitioners and also improve overall eDNA data quality.

Predator–prey interactions are critical in understanding how communities function. However, we need to describe intraspecific variation in diet to accurately depict those interactions. Harbor seals (Phoca vitulina) are an abundant marine predator that prey on species of conservation concern. We estimated intrapopulation feeding diversity (variation in feeding habits between individuals of the same species) of harbor seals in the Salish Sea. Estimates of feeding diversity were examined relative to sex, month, and location using a novel approach that combined molecular techniques, repeated cross‐sectional sampling of scat, and a specialization metric (within‐individual consistency in diet measured by the Proportional Similarity Index (PSi)). Based on 1,083 scat samples collected from five haul‐out sites during four nonsequential years, we quantified diet using metabarcoding techniques and determined the sex of the scat depositor using a molecular assay. Results suggest that intrapopulation feeding diversity was present. Specialization was high over short periods (24–48 hr, PSi = 0.392, 95% CI = 0.013, R = 100,000) and variable in time and space. Females showed more specialization than males, particularly during summer and fall. Additionally, demersal and benthic prey species were correlated with more specialized diets. The latter finding suggests that this type of prey likely requires specific foraging strategies and that there are trade‐offs between pelagic and benthic foraging styles for harbor seals. This differential feeding on prey species, as well as between sexes of harbor seals, indicates that predator–prey interactions in harbor seals are complex and that each sex may have a different impact on species of conservation concern. As such, describing intrapopulation feeding diversity may unravel hitherto unknown complex predator–prey interactions in the community., We estimated intrapopulation feeding diversity of harbor seals in the Salish Sea relative to sex, time, and location with a novel approach that combined molecular techniques, repeated cross‐sectional sampling of scat, and a specialization metric (within‐individual consistency in diet). Differential feeding on prey species was revealed, as well as differences between sexes of harbor seals. This indicates that predator–prey interactions in harbor seals are complex and that each sex may have a different impact on species of conservation concern. As such, describing intraspecific variation in diet may unravel hitherto unknown complex predator–prey interactions in the community.

Sex-biased predation, a predator’s bias for one prey sex over the other, can have important demographic impacts on prey species of conservation concern. Yet, it is difficult to measure in the wild. Molecular scatology has been used to indirectly determine the proportion of prey items consumed in the diet, and it may be possible to apply similar approaches to determine the sex-biased consumption of prey items. We developed a molecular method to indirectly examine sex-specific predation employing scat, focusing on predator–prey interactions between Chinook salmon (Oncorhynchus tshawytscha) and harbor seals (Phoca vitulina richardii). We established that the proportions of male and female Chinook DNA can be determined in a controlled sample by measuring a Y-linked marker, growth hormone pseudogene, using qPCR. We then applied the assay to harbor seal scat samples. Although the assay amplified in 83% of scat samples, 27% of samples quantified had an estimated male proportion > 1, which may have been due to a lack of robustness of the PCR assay in samples. Lastly, we constructed a biomass calibration curve to determine whether DNA measurements could estimate the proportions of male and female biomass consumed. The calibration curve was skewed by high male DNA density precluding our ability to quantify the relative amounts. We demonstrated that nuclear prey markers can be amplified in predator scat, however, contamination and extreme DNA density differences between the prey sexes may pose practical difficulties to estimate the relative amounts of male to female biomass consumed.

Environmental DNA (eDNA) detection of aquatic invasive species is currently at the forefront of aquatic conservation efforts because the methodology provides a cost effective and sensitive means to detect animals at low densities. Developments in eDNA technologies have improved detection probabilities for rare, indicator, and invasive species over the past decade. However, standard lab analysis can take days or weeks before results are available and is prohibitive when rapid management decisions are required for mitigation. Here, we investigated the performance of a real-time quantitative PCR system for on-site eDNA detection of New Zealand mud snails (Potamopyrgus antipodarum). Six sites in western Washington, USA were sampled using the rapid eDNA technique and traditional methods, with five samples per site. On-site eDNA detection of mud snails resulted in a 10% increase in positive sites (16/30 = 53% positive) relative to visual surveys (13/30 = 43% positive). In addition, positive associations were observed between mud snail eDNA concentration (eDNA copies per reaction) and the number of mud snail individuals at each site (R2 = 0.78). We show that the rapid on-site eDNA technology can be effective for detection and quantification of New Zealand mud snails in freshwaters. This on-site eDNA detection approach could possibly be used to initiate management protocols that allow for more rapid responses during the onset of biological invasions.

Many marine mammal predators, particularly pinnipeds, have increased in abundance in recent decades, generating new challenges for balancing human uses with recovery goals via ecosystem-based management. We used a spatio-temporal bioenergetics model of the Northeast Pacific Ocean to quantify how predation by three species of pinnipeds and killer whales (Orcinus orca) on Chinook salmon (Oncorhynchus tshawytscha) has changed since the 1970s along the west coast of North America, and compare these estimates to salmon fisheries. We find that from 1975 to 2015, biomass of Chinook salmon consumed by pinnipeds and killer whales increased from 6,100 to 15,200 metric tons (from 5 to 31.5 million individual salmon). Though there is variation across the regions in our model, overall, killer whales consume the largest biomass of Chinook salmon, but harbor seals (Phoca vitulina) consume the largest number of individuals. The decrease in adult Chinook salmon harvest from 1975–2015 was 16,400 to 9,600 metric tons. Thus, Chinook salmon removals (harvest + consumption) increased in the past 40 years despite catch reductions by fisheries, due to consumption by recovering pinnipeds and endangered killer whales. Long-term management strategies for Chinook salmon will need to consider potential conflicts between rebounding predators or endangered predators and prey.

Conflicts can arise when the recovery of one protected species limits the recovery of another through competition or predation. The recovery of many marine mammal populations on the west coast of the United States has been viewed as a success; however, within Puget Sound in Washington State, the increased abundance of three protected pinniped species may be adversely affecting the recovery of threatened Chinook salmon (Oncorhynchus tshawytscha) and endangered killer whales (Orcinus orca) within the region. Between 1970 and 2015, we estimate that the annual biomass of Chinook salmon consumed by pinnipeds has increased from 68 to 625 metric tons. Converting juvenile Chinook salmon into adult equivalents, we found that by 2015, pinnipeds consumed double that of resident killer whales and six times greater than the combined commercial and recreational catches. We demonstrate the importance of interspecific interactions when evaluating species recovery. As more protected species respond positively to recovery efforts, managers should attempt to evaluate tradeoffs between these recovery efforts and the unintended ecosystem consequences of predation and competition on other protected species.

Knowing the species and life stages of prey that predators consume is important for understanding the impacts that predation may have on prey populations, but traditional methods for determining diets often cannot provide sufficient detail. We combined data from two methods of scat analysis (DNA metabarcoding and morphological prey ID) to quantify the species and life stages of salmon (Oncorhynchus spp.) consumed by harbour seals (Phoca vitulina) in the Strait of Georgia, Canada, where juvenile Chinook (Oncorhynchus tshawytscha) and coho (Oncorhynchus kisutch) salmon survival is poor. Harbour seals primarily consumed adult salmon of lesser conservation concern in the fall (August–November): chum (Oncorhynchus keta: 18.4%), pink (Oncorhynchus gorbuscha: 12.6%), sockeye (Oncorhynchus nerka: 7.4%), Chinook (7.1%), and coho (1.8%). However, the opposite species trend occurred during the spring when seals preferred juvenile salmon of greater conservation concern (April–July): coho (2.9%), Chinook (2.9%), sockeye (2.5%), pink (1.4%), and chum (0.8%) — percentages that can equate to many individuals consumed. Our data suggest that harbour seals select juveniles of salmon species that out-migrate at ages >1 year and provide evidence of a potential causal relationship between harbour seal predation and juvenile salmon survival trends.

Advances in technology are making it easier for rapid field detection of microbes in aquaculture. Specifically, real-time quantitative PCR (qPCR) analysis, which has traditionally been confined to laboratory-based protocols, is now available in a handheld, field-portable system. The feasibility of using the Biomeme handheld qPCR system for rapid (

Advances in DNA sequencing technology have revolutionized the field of molecular analysis of trophic interactions, and it is now possible to recover counts of food DNA sequences from a wide range of dietary samples. But what do these counts mean? To obtain an accurate estimate of a consumer's diet should we work strictly with data sets summarizing frequency of occurrence of different food taxa, or is it possible to use relative number of sequences? Both approaches are applied to obtain semi‐quantitative diet summaries, but occurrence data are often promoted as a more conservative and reliable option due to taxa‐specific biases in recovery of sequences. We explore representative dietary metabarcoding data sets and point out that diet summaries based on occurrence data often overestimate the importance of food consumed in small quantities (potentially including low‐level contaminants) and are sensitive to the count threshold used to define an occurrence. Our simulations indicate that using relative read abundance (RRA) information often provides a more accurate view of population‐level diet even with moderate recovery biases incorporated; however, RRA summaries are sensitive to recovery biases impacting common diet taxa. Both approaches are more accurate when the mean number of food taxa in samples is small. The ideas presented here highlight the need to consider all sources of bias and to justify the methods used to interpret count data in dietary metabarcoding studies. We encourage researchers to continue addressing methodological challenges and acknowledge unanswered questions to help spur future investigations in this rapidly developing area of research.

Sex‐specific diet information is important in the determination of predator impacts on prey populations. Unfortunately, the diet of males and females can be difficult to describe, particularly when they are marine predators. We combined two molecular techniques to describe haul‐out use and prey preferences of male and female harbor seals (Phoca vitulina) from Comox and Cowichan Bay (Canada) during 2012–2013. DNA metabarcoding quantified the diet proportions comprised of prey species in harbor seal scat, and qPCR determined the sex of the individual that deposited each scat. Using 287 female and 260 male samples, we compared the monthly sex ratio with GLMs and analyzed prey consumption relative to sex, season, site, and year with PERMANOVA. The sex ratio between monthly samples differed widely in both years (range = 12%–79% males) and showed different patterns at each haul‐out site. Male and female diet differed across both years and sites: Females consumed a high proportion of demersal fish species while males consumed more salmonid species. Diet composition was related to both sex and season (PERMANOVA: R 2 = 27%, p

DNA metabarcoding is a powerful new tool allowing characterization of species assemblages using high-throughput amplicon sequencing. The utility of DNA metabarcoding for quantifying relative species abundances is currently limited by both biological and technical biases which influence sequence read counts. We tested the idea of sequencing 50/50 mixtures of target species and a control species in order to generate relative correction factors (RCFs) that account for multiple sources of bias and are applicable to field studies. RCFs will be most effective if they are not affected by input mass ratio or co-occurring species. In a model experiment involving three target fish species and a fixed control, we found RCFs did vary with input ratio but in a consistent fashion, and that 50/50 RCFs applied to DNA sequence counts from various mixtures of the target species still greatly improved relative abundance estimates (e.g. average per species error of 19 ± 8% for uncorrected vs. 3 ± 1% for corrected estimates). To demonstrate the use of correction factors in a field setting, we calculated 50/50 RCFs for 18 harbour seal (Phoca vitulina) prey species (RCFs ranging from 0.68 to 3.68). Applying these corrections to field-collected seal scats affected species percentages from individual samples (Δ 6.7 ± 6.6%) more than population-level species estimates (Δ 1.7 ± 1.2%). Our results indicate that the 50/50 RCF approach is an effective tool for evaluating and correcting biases in DNA metabarcoding studies. The decision to apply correction factors will be influenced by the feasibility of creating tissue mixtures for the target species, and the level of accuracy needed to meet research objectives.

International audience; Accurately estimating predators’ diets at relevant spatial and temporal scales is key to understanding animals’ energetics and fitness, particularly in populations whose decline might be related to their diet such as northern fur seals Callorhinus ursinus. Our goals were to improve the accuracy of diet estimates and extend understanding of feeding ecology by combining 2 scat-based methods of diet determination (hard-part identification and DNA-metabarcoding) with stable isotope measurements and individual behavioural data. We collected 98 scats on a northern fur seal breeding colony. We also tracked 20 females with biologgers, and took blood samples to determine δ13C and δ15N values as proxies for seal foraging habitat and diet. Results show that diet composition from hard-parts analysis corresponded well with DNA results, with DNA yielding a greater diversity of prey species at a finer taxonomic level. Overall, scat-based methods showed that seals mostly fed on neritic shelf-associated prey. Cluster analyses of combined hard-parts and DNA results however identified 2 diet groups, one mostly neritic and the other mostly pelagic. Stable isotopes and behavioural data revealed that 40% of seals fed in oceanic waters on pelagic prey. This is more than indicated by scat-based analyses, which are likely biased towards animals foraging closest to the colony and underestimate some dietary specializations within the population. Consequently, the combination of multiple methods for diet identification with at-sea tracking of individuals can help identify and quantify specialist groups within a population and provide a wider spatial and temporal ecological context for dietary analysis.

Brucella ceti and Brucella pinnipedialis have been documented as occurring in marine mammals, and B. ceti has been identified in 3 naturally acquired human cases. Seroconversion and infection patterns in Pacific Northwest harbor seals ( Phoca vitulina richardii) and North Atlantic hooded seals ( Cystophora cristata) indicate post-weaning exposure through prey consumption or lungworm infection, suggesting fish and possibly invertebrates play an epizootiologic role in marine Brucella transmission and possible foodborne risk to humans. We determined if real-time quantitative PCR (qPCR) assays can detect marine Brucella DNA in fish DNA. Insertion sequence (IS) 711 gene and sequence type (ST)27 primer–probe sets were used to detect Brucella associated with marine mammals and human zoonotic infections, respectively. First, DNA extracts from paired-species fish (containing 2 species) samples were tested and determined to be Brucella DNA negative using both IS 711 and ST27 primer–probe sets. A representative paired-species fish DNA sample was spiked with decreasing concentrations of B. pinnipedialis DNA to verify Brucella detection by the IS 711 primer–probe within fish DNA. A standard curve, developed using isolated DNA from B. pinnipedialis, determined the limit of detection. Finally, the IS 711 primer–probe was used to test Atlantic cod ( Gadus morhua) DNA extracts experimentally infected with the B. pinnipedialis hooded seal strain. In culture-positive cod tissue, the IS 711 limit of detection was ~1 genome copy of Brucella. Agreement between culture and PCR results for the 9 positive and 9 negative cod tissues was 100%. Although a larger sample set is required for validation, our study shows that qPCR can detect marine Brucella in fish.

A goal of many environmental DNA barcoding studies is to infer quantitative information about relative abundances of different taxa based on sequence read proportions generated by high-throughput sequencing. However, potential biases associated with this approach are only beginning to be examined. We sequenced DNA amplified from faeces (scats) of captive harbour seals (Phoca vitulina) to investigate whether sequence counts could be used to quantify the seals' diet. Seals were fed fish in fixed proportions, a chordate-specific mitochondrial 16S marker was amplified from scat DNA and amplicons sequenced using an Ion Torrent PGM™. For a given set of bioinformatic parameters, there was generally low variability between scat samples in proportions of prey species sequences recovered. However, proportions varied substantially depending on sequencing direction, level of quality filtering (due to differences in sequence quality between species) and minimum read length considered. Short primer tags used to identify individual samples also influenced species proportions. In addition, there were complex interactions between factors; for example, the effect of quality filtering was influenced by the primer tag and sequencing direction. Resequencing of a subset of samples revealed some, but not all, biases were consistent between runs. Less stringent data filtering (based on quality scores or read length) generally produced more consistent proportional data, but overall proportions of sequences were very different than dietary mass proportions, indicating additional technical or biological biases are present. Our findings highlight that quantitative interpretations of sequence proportions generated via high-throughput sequencing will require careful experimental design and thoughtful data analysis.

The Great Shearwater (Puffinus gravis) is an abundant pelagic seabird that undertakes transequatorial migrations between the North and South Atlantic Ocean. This species is a useful indicator of large-scale alterations in marine dynamics due to its wide geographic range, long-distance migrations, and relative abundance. From 1993 to 2011, 12 separate mortality events, with 4,961 Great Shearwaters recovered, were documented along the eastern coast of the United States. Of these, seven events (n=4,885) occurred in the Southeast (SE) and five (n=76) in the Northeast (NE) United States. The cause of death was determined either by necropsy (n=60) or external examination (n=4,901). All Great Shearwaters stranded along the SE United States were emaciated while 58% were emaciated in the NE United States. No plastic was observed in Great Shearwaters in the SE US (n=27), but the gastrointestinal tract of 82% (n=27) of all stranded birds along the NE United States had at least one plastic bead. There was no evidence of infectious disease or heavy metals in stranded Great Shearwaters examined (n=14, from the 2005 SE event). Stable isotope analysis of feathers (n=9, from a 2007 SE event) suggests dietary differences between emaciated stranded birds and live-caught healthy birds. The temporal distribution of stranding detections suggests a general increase in the number of observed Great Shearwater strandings over the past two decades. From 1993 to 2000 there were a total of three mortality events with 296 individual Great Shearwaters. However, there was a threefold increase in the number of mortality events from 2001 to 2011 (nine events involving 4,665 individuals). The causes of this apparent increase in strandings are unknown but may be due to an increase in reporting effort over the past two decades combined with changing oceanographic conditions in the South Atlantic Ocean, leading to large-scale mortality of emaciated Great Shearwaters along the east coast of the United States.

Harbor seal (Phoca vitulina richardii) populations in the inland waters of Washington and British Columbia are at or near carrying capacity. Stranded pups often are collected and admitted to rehabilitation centers, and then released when they reach a weight of 22 kg and meet a variety of preestablished health and release conditions. While rehabilitation is common practice, it is unclear if rehabilitated seal pups behave like wild weaned pups. Using satellite transmitters, we compared movement patterns of 10 rehabilitated pups with 10 wild weaned pups. When released, rehabilitated seals were longer and heavier than wild pups, while wild pups had a larger mean axillary girth. No clinically different blood parameters were detected. On average, rehabilitated harbor seal pups traveled nearly twice as far cumulatively, almost three times as far daily, and dispersed over three times as far from the release site compared to wild weaned seals. Additionally, wild harbor seals transmitted nearly twice as long as did rehabilitated seals. These patterns suggest that learned behavior during the brief 3–4 wk nursing period likely enables wild harbor seal pups to move less daily and remain closer to their weaning site than rehabilitated pups.

Sharks are of commercial, research, conservation, and exhibition importance but we know little regarding health parameters and population status for many species. Here we present health indicators and species comparisons for adults of three common wild-caught species: 30 Atlantic sharpnose sharks (Rhizoprionodon terraenovae) and 31 bonnethead sharks (Sphyrna tiburo) from the western Atlantic, and 30 spiny dogfish sharks (Squalus acanthias) from the eastern Pacific. All animals were captured during June-July 2009 and 2010. Median values and preliminary reference intervals were calculated for hematology, plasma biochemistry, trace nutrients, and vitamin A, E, and D concentrations. Significant differences, attributable to physiologic differences among the species, were found in the basic hematologic and plasma biochemistry variables. Significant species differences in arsenic and selenium plasma concentrations were found and appear to coincide with diet and habitat variability among these three species. Vitamin E was significantly higher in the bonnethead shark, again related to the foraging ecology and ingestion of plant material by this species. The Atlantic sharpnose had significantly higher vitamin A concentrations, supported by the higher proportion of teleosts in the diet. Vitamin D was below the limit of quantification in all three species. These preliminary reference intervals for health variables can be used to assess and monitor the population health and serve as indicators of nutritional status in these populations of wild elasmobranchs.

Ecologists are increasingly interested in quantifying consumer diets based on food DNA in dietary samples and high-throughput sequencing of marker genes. It is tempting to assume that food DNA sequence proportions recovered from diet samples are representative of consumer's diet proportions, despite the fact that captive feeding studies do not support that assumption. Here, we examine the idea of sequencing control materials of known composition along with dietary samples in order to correct for technical biases introduced during amplicon sequencing and biological biases such as variable gene copy number. Using the Ion Torrent PGM(©) , we sequenced prey DNA amplified from scats of captive harbour seals (Phoca vitulina) fed a constant diet including three fish species in known proportions. Alongside, we sequenced a prey tissue mix matching the seals' diet to generate tissue correction factors (TCFs). TCFs improved the diet estimates (based on sequence proportions) for all species and reduced the average estimate error from 28 ± 15% (uncorrected) to 14 ± 9% (TCF-corrected). The experimental design also allowed us to infer the magnitude of prey-specific digestion biases and calculate digestion correction factors (DCFs). The DCFs were compared with possible proxies for differential digestion (e.g. fish protein%, fish lipid%) revealing a strong relationship between the DCFs and percent lipid of the fish prey, suggesting prey-specific corrections based on lipid content would produce accurate diet estimates in this study system. These findings demonstrate the value of parallel sequencing of food tissue mixtures in diet studies and offer new directions for future research in quantitative DNA diet analysis.