Sex-specific diet differences in harbor seals (Phoca vitulina) via spatial assortment.

The lack of recovery of Chinook salmon (Oncorhynchus tshawytscha) in the Pacific Northwest has been blamed in part on predation by pinnipeds, particularly the harbor seal (Phoca vitulina). Previous work at a limited number of locations has shown that male seal diet contains more salmon than that of female seals and that sex ratios at haul-out sites differ spatiotemporally. This intrapopulation variation in predation may result in greater effects on salmon than suggested by models assuming equal spatial distribution and diet proportion. To address the generality of these patterns, we examined the sex ratios and diet of male and female harbor seals from 13 haul-out sites in the inland waters of Washington State and the province of British Columbia during 2012-2018. DNA metabarcoding was conducted to determine prey species proportions of individual scat samples. The sex of harbor seals was then determined from each scat matrix sample with the use of quantitative polymerase chain reaction (qPCR). We analyzed 2405 harbor seal scat samples using generalized linear mixed models (GLMMs) to examine the factors influencing harbor seal sex ratio at haul-out sites and permutational multivariate analysis of variance (PERMANOVA) to examine the influence of sex and haul-out site on harbor seal diet composition. We found that the overall sex ratio was 1:1.02 (female:male) with notable spatiotemporal variation. Salmoniformes were about 2.6 times more abundant in the diet of males than in the diet of females, and Chinook salmon comprised ca. three times more of the average male harbor seal's diet than the average female's diet. Based on site-specific sex ratios and diet data, we identified three haul-out sites where Chinook salmon appear to be under high predation pressure by male harbor seals: Cowichan Bay, Cutts Area, and Fraser River. Our study indicates that combining sex-specific pinniped diet data with the sex ratio of haul-out sites can help identify priority sites of conservation concern.

Metabarcoding analysis provides insight into the link between prey and plant intake in a large alpine cat carnivore, the snow leopard.

Species of the family Felidae are thought to be obligate carnivores. However, detection of plants in their faeces raises questions about the role of plants in their diet. This is particularly true for the snow leopard (Panthera uncia). Our study aimed to comprehensively identify the prey and plants consumed by snow leopards. We applied DNA metabarcoding methods on 90 faecal samples of snow leopards collected in Kyrgyzstan, employing one vertebrate and four plant markers. We found that argali (Ovis ammon) was detected only from male snow leopards. Myricaraia sp. was the most consumed among 77 plant operational taxonomic units found in snow leopard samples. It frequently appeared in samples lacking any prey animal DNA, indicating that snow leopards might have consumed this plant especially when their digestive tracts were empty. We also observed differences in the patterns of plant consumption between male and female snow leopards. Our comprehensive overview of prey and plants detected in the faeces of snow leopards and other sympatric mammals will help in formulating hypotheses and guiding future research to understand the adaptive significance of plant-eating behaviour in felids. This knowledge supports the enhancement of their captive environments and the conservation planning of their natural habitats.

Molecular analyses of carangid fish diets reveal inter-predation, dietary overlap, and the importance of early life stages in trophic ecology.

Carangid fishes are commercially important in fisheries and aquaculture. They are distributed worldwide in both tropical and subtropical marine ecosystems. Their role in food webs is often unclear since their diet cannot be easily identified by traditional gut content analysis. They are suspected to prey on pelagic and benthic species, with clupeiform fishes being important dietary items for some species, though it is unknown whether carangids share food resources or show trophic segregation. Here, we used metabarcoding to overcome traditional challenges of taxonomic approaches to analyze the diet of seven carangid species caught as bycatch in the Brazilian southwest Atlantic sardine fishery. Stomach contents were processed from the following species: Caranx crysos, Caranx latus, Chloroscombrus chrysurus, Hemicaranx amblyrhynchus, Oligoplites saliens, Selene setapinnis, and Trachinotus carolinus. Identified diets were dominated by teleost fishes. The C. latus diet was the most distinct among the seven species, preferentially consuming Engraulis anchoita, but H. amblyrhynchus, O. saliens, and S. setapinnis also showed a trend of predominantly consuming small pelagic fishes. Finally, we found evidence of inter-predation in carangids, especially strong between S. setapinnis and C. crysos, suggesting that consumption of early life stages may result in indirect competition through reduced recruitment in these fishes. These findings provide unprecedented insights into the biodiversity in marine ecosystems, especially the poorly known diet of carangid fishes.

Transitioning to a Plant-Based Menu in Childcare: Identifying the Nutritional, Financial, and Logistical Considerations.

International health organizations have called for a shift towards more plant-based foods as a way of promoting both individual health and environmental sustainability. Given the high percentage of children in Canada who attend childcare and the high volume of food provided in childcare, transitioning menus to incorporate plant-based foods could have important implications for both planetary and child health. The purpose of this case study is to describe a childcare centre's transition to a plant-based menu. A detailed nutritional analysis of the menu was conducted. The financial and logistical implications of the transitions to a plant-based menu were also assessed. Nutritional analysis revealed that the plant-based menu met or exceeded the daily nutrient requirement for all the key nutrients explored. Financially, the transition led to a 9% reduction in food costs. Logistically, the transition led to improved efficiency and safety with regard to food preparation, with substantially fewer tailored meals due to allergies and dietary restrictions required after the transition. These novel findings are relevant for food service administrators interested in transitioning to a plant-based menu as well as public health dietitians who could support the transition.

Prey ration, temperature, and predator species influence digestion rates of prey DNA inferred from qPCR and metabarcoding.

Diet analysis is a vital tool for understanding trophic interactions and is frequently used to inform conservation and management. Molecular approaches can identify diet items that are impossible to distinguish using more traditional visual-based methods. Yet, our understanding of how different variables, such as predator species or prey ration size, influence molecular diet analysis is still incomplete. Here, we conducted a large feeding trial to assess the impact that ration size, predator species, and temperature had on digestion rates estimated with visual identification, qPCR, and metabarcoding. Our trial was conducted by feeding two rations of Chinook salmon (Oncorhynchus tshawytscha) to two piscivorous fish species (largemouth bass [Micropterus salmoides] and channel catfish [Ictalurus punctatus]) held at two different temperatures (15.5 and 18.5°C) and sacrificed at regular intervals up to 120 h from the time of ingestion to quantify the prey contents remaining in the digestive tract. We found that ration size, temperature, and predator species all influenced digestion rate, with some indication that ration size had the largest influence. DNA-based analyses were able to identify salmon smolt prey in predator gut samples for much longer than visual analysis (~12 h for visual analysis vs. ~72 h for molecular analyses). Our study provides evidence that modelling the persistence of prey DNA in predator guts for molecular diet analyses may be feasible using a small set of controlling variables for many fish systems.

Metabarcoding of fecal DNA reveals the broad and flexible diet of a globally endangered bird.

Knowing the diet of endangered wild animals is a prerequisite for species-specific conservation and habitat management. The Sichuan partridge Arborophila rufipectus is a globally endangered Galliformes species endemic to the mountains of southwest China. Existing information on the diet of this species is biased and fragmented owing to traditional observation methods. Little is known about their dietary composition or how they respond to temporal variations in food resources throughout the year. In this study, a dietary analysis was performed on 60 fecal samples using DNA Metabarcoding of invertebrates and plants to determine the primary animal and plant components of the diet across 3 critical periods of adult life history (breeding, postbreeding wandering, and overwintering). Preys from the dipteran order, followed by the lepidopteran and araneaen spp., were the predominant, animal-derived foods. Symplocos, Rubus, Celastrus, Holboellia, and Actinidia spp. supply a large abundance of fruits and seeds for this omnivorous bird. Substantial temporal dietary changes among the 3 periods and a general shift toward lower dietary diversity during the breeding season were observed, suggesting that the Sichuan partridge can adjust their diet according to the availability of food resources and their own needs. Characterizing the composition and seasonal changes in Sichuan partridge diets informs the habitat management of native flora (the plant taxa that can generate berries and seeds, such as Symplocos, Rubus, Celastrus, and Holboellia, which are likely of conservation interest) to achieve full life-cycle conservation.

Diet composition of wild columbiform birds: next-generation sequencing of plant and metazoan DNA in faecal samples.

Accurate knowledge of a species' diets is fundamental to understand their ecological requirements. Next-generation sequencing technology has become a powerful and non-invasive tool for diet reconstruction through DNA metabarcoding. Here, we applied those methods on faecal samples of Common Woodpigeons Columba palumbus, European Turtle Doves Streptopelia turtur, and Stock Doves C. oenas to investigate their dietary composition. By applying primer pairs targeting both the ITS2 region of plant nuclear DNA and the mitochondrial COI region of metazoan DNA, we provide a complete picture of the food ingested and estimate the dietary overlap between the columbiform species during the breeding season. Animal DNA was present very rarely, and a diverse range of plants from the class Spermatopsida dominated the diet, with Asteraceae, Brassicaceae, Cucurbitaceae, Fabaceae, and Poaceae as the most frequently represented families. Generally, we detected a variability between species but also amongst individual samples. Plant species already known from previous studies, mainly visual analyses, could be confirmed for our individuals sampled in Germany and the Netherlands. Our molecular approach revealed new plant taxa, e.g. plants of the families Malvaceae for Woodpigeons, Lythraceae for Turtle Doves, and Pinaceae for Stock Doves, not found in previous studies using visual analyses. Although most of the plant species observed were of wild origin, the majority of cultivated plants found were present in higher frequencies of occurrence, suggesting that cultivated food items likely constitute an important part of the diet of the studied species. For Turtle Doves, a comparison with previous studies suggested regional differences, and that food items (historically) considered as important part of their diet, such as Fumitory Fumaria sp. and Chickweed Stellaria media, were missing in our samples. This indicates that regional variations as well as historic and current data on diet should be considered to plan tailored seed mixtures, which are currently proposed as an important management measure for conservation of the rapidly declining Turtle Dove.

Diet of a threatened rattlesnake (eastern massasauga) revealed by DNA metabarcoding.

Characterizing the diet of imperiled species using minimally invasive methods is crucial to understanding their ecology and conservation requirements. Here, we apply a DNA metabarcoding approach to study the diet of the eastern massasauga rattlesnake (Sistrurus catenatus), a Federally Threatened snake found throughout the Great Lakes region. Eighty-three fecal samples collected across 10 different massasauga populations located in Michigan, USA, were sequenced, with 70 samples containing prey DNA. We used universal metazoan primers and developed a host-specific oligonucleotide blocker to characterize their diet. We identified at least 12 different prey species, with eastern massasaugas exhibiting opportunistic feeding and a strong preference towards small mammals. Meadow voles (Microtus pennsylvanicus) were the most common prey item (70% of diet) followed by the northern short-tailed shrew (Blarina brevicauda) and masked shrew (Sorex cinereus; 15.7% of diet each), along with occasional bird and snake prey. Adult individuals exhibited a more generalized diet, consuming a larger number of prey taxa on average. Younger snakes consumed a smaller variety of prey items and tended to consume smaller-sized mammals such as masked shrews (Sorex cinereus) and northern short-tailed shrews (Blarina brevicauda). We conclude that small mammals are a crucial part of eastern massasauga rattlesnake diet and recommend this be taken into consideration when conservation strategies are developed. The methods developed in this study can be applied to other reptile species, providing an accurate, minimally invasive, and thorough diet assessment for at-risk reptile species.

DNA-based seed intake quantification for enhanced ecological risk assessment of small mammals.

To prevent the non-acceptable effects of agrochemicals on arable fields, Environmental Risk Assessment (ERA) aims to assess and protect against a wide range of risks due to stressors to non-target species. While exposure to stress is a key factor in ERA models, exposure values are difficult to obtain and rely on laboratory studies with often debatable relevance to field situations. To improve intake estimates, data from realistic field-based scenarios are needed. We developed calibration curves relating known seed numbers of up to 20 onion and carrot seeds consumed by wild-caught wood mice (Apodemus sylvaticus) to the seed DNA content in the feces. Based on these inferred quantitative relationships, a field trial was run to determine seed intake in a natural setting using realistic levels of seed spillage. Onion DNA was detected in the fecal samples of the wood mice caught in the field, which resembled a seed intake of up to 1 onion seed. No intake of carrot seeds was detected. This is the first-ever study to quantify seed intake in a realistic field scenario using a DNA-based analysis, showing that accurate seed intake estimates can be obtained. Our approach can help to improve risk assessment models through its minimally-invasive and accurate assessment of seed intake by ERA representative and non-target species, which would otherwise be undetectable with traditional methods. Our novel approach and its results are highly relevant to studies of food intake and diet composition for basic and applied research alike.

Toward absolute abundance for conservation applications: Estimating the number of contributors via microhaplotype genotyping of mixed-DNA samples.

Molecular methods including metabarcoding and quantitative polymerase chain reaction have shown promise for estimating species abundance by quantifying the concentration of genetic material in field samples. However, the relationship between specimen abundance and detectable concentrations of genetic material is often variable in practice. DNA mixture analysis represents an alternative approach to quantify specimen abundance based on the presence of unique alleles in a sample. The DNA mixture approach provides novel opportunities to inform ecology and conservation by estimating the absolute abundance of target taxa through molecular methods; yet, the challenges associated with genotyping many highly variable markers in mixed-DNA samples have prevented its widespread use. To advance molecular approaches for abundance estimation, we explored the utility of microhaplotypes for DNA mixture analysis by applying a 125-marker panel to 1179 Chinook salmon (Oncorhynchus tshawytscha) smolts from the Sacramento-San Joaquin Delta, California, USA. We assessed the accuracy of DNA mixture analysis through a combination of mock mixtures containing DNA from up to 20 smolts and a trophic ecological application enumerating smolts in predator diets. Mock DNA mixtures of up to 10 smolts could reliably be resolved using microhaplotypes, and increasing the panel size would likely facilitate the identification of more individuals. However, while analysis of predator gastrointestinal tract contents indicated DNA mixture analysis could discern the presence of multiple prey items, poor and variable DNA quality prevented accurate genotyping and abundance estimation. Our results indicate that DNA mixture analysis can perform well with high-quality DNA, but methodological improvements in genotyping degraded DNA are necessary before this approach can be used on marginal-quality samples.

Estimating predation rates from molecular gut content analysis.

Several methods have been published to estimate per capita predation rates from molecular gut content analysis relying on intuitive understanding of predation, but none have been formally derived. We provide a theoretical framework for estimating predation rates to identify an accurate method and lay bare its assumptions. Per capita predation can be estimated by multiplying the prey decay rate and the prey quantity in the predators. This assumes that variation in per capita predation rate is approximately normally distributed, prey decay occurs exponentially, and predation is in steady state. We described several ways to estimate steady state predation, including using only qualitative presence-absence data to estimate the decay rate and in addition, we provided a method for estimating per capita predation rate when predation is not in steady state. We used previously published data on aphid consumption by a ladybird beetle in a feeding trial to calculate the predation rate and compare published methods with this theoretically derived method. The estimated predation rate (3.29 ± 0.27 aphids/h) using our derived method was not significantly different from the actual predation rate, 3.11 aphids/h. In contrast, previously published methods were less accurate, underestimating the predation rate (0.33 ± 0.02 to 1.66 ± 0.8 aphids/h) or overestimating it (3.64 ± 0.30 aphids/h). In summary, we provide methods to estimate predation rates even when variation in predation rates is not exactly normally distributed and not in steady state and demonstrate that the prey decay rate, and not the prey detection period, is required.

Diet of Free Ranging American Mink (Neovison vison) in Denmark.

Non-native American mink (Neovison vison) is a widely spread predator in Denmark. The feral population in mainland Denmark consists of captive-born mink that escaped from fur farms and wild mink born in nature, whereas the population on Bornholm is almost all wild-born mink. In this study, the diets of feral mink in mainland Denmark and on the island of Bornholm are analyzed. The aim of this study was to examine (1) whether the diet of the larger captive-born mink differs from that of the smaller wild-born mink, (2) assess the regional variations between the diets of mink in mainland Denmark and on Bornholm, and (3) investigate the seasonal variation in the diet composition of mink. The stomach contents of 364 mink (243 wild-born and 114 captive-born) culled in the years 2019-2022 were analyzed. Of these, 203 mink were from mainland Denmark, and 154 were from Bornholm. No significant differences were found between the diets of captive-born mink and wild-born mink or the mink found in mainland Denmark and on the island of Bornholm. Significantly more empty stomachs were found during spring than during the summer and autumn, suggesting a bottleneck in the diet during spring.

Diet of the Insular Lizard, Podarcis lilfordi (Günther, 1874): Complementary Morphological and Molecular Approaches.

The diets of insular lizards are extremely varied, depending on the different environmental characteristics of each island population. This is particularly evident in the case of the populations of small coastal islets of the Balearic Islands, where the Balearic lizard, Podarcis lilfordi, is found. The study of trophic ecology carried out by means of traditional tools, such as morphological analysis of feces, has made it possible to detect numerous prey and nutritional elements. However, these methods are clearly insufficient, as some rare groups are not detected. It is also difficult to identify remains of marine subsidies or of foods contributed to these small islands by other predators, such as seabirds. The current study demonstrates the advantages of combining morphological diet analysis with the molecular study of individual feces samples obtained from the same populations. We obtained a greater diversity of prey groups using the combined methodologies, with each method identifying prey items that were not detected using the other method. Particularly, the study of diets at the molecular level identified plant species consumed by lizards that were, occasionally, not identified in morphological analyses. Conversely, the traditional morphological study of an equivalent number of fecal samples allowed for the identification of several prey groups that had not been detected in the molecular study. From this viewpoint, the advantages and disadvantages of each methodology are discussed.

Food webs reveal coexistence mechanisms and community organization in carnivores.

Globally, massive carnivore guild extirpations have led to trophic downgrading and compromised ecosystem services. However, the complexity of multi-carnivore food webs complicates accurate identification of species interactions and community organization. Here, we used fecal DNA metabarcoding to investigate three communities that together encompass eight large- and meso-carnivore species and their 44 prey taxa of the Qinghai-Tibet Plateau (QTP), one of the last places on Earth that still harbors intact carnivore assemblages. Quantitative food-web analyses revealed pronounced interspecific variations in the carnivores' prey compositions and dietary partitioning both between and within guilds. Additionally, body masses of the carnivores and their prey exhibited consistent hump-shaped correlations across communities. Overall, differences in prey diversity, size category, and proportional utilization among the carnivore species result in trophic niche segregation that likely promotes carnivore coexistence in the harsh QTP environment. Network structure analyses detected significant modularity in all food webs but nestedness in only one. Furthermore, network characterization identified pikas (Ochotona spp.), bharal (Pseudois nayaur), and domestic yak (Bos grunniens) as potential keystone prey across the areas. Our results paint a holistic and detailed picture of the QTP carnivore assemblages' trophic networks and demonstrate that the combined use of the molecular dietary approach and network analysis can generate structural insights into carnivore coexistence and can identify functionally important species in complex communities. Such knowledge can help safeguard carnivore guild integrity and enhance community resilience to environmental perturbations in the sensitive QTP ecosystems.

Toward quantitative metabarcoding.

Amplicon-sequence data from environmental DNA (eDNA) and microbiome studies provide important information for ecology, conservation, management, and health. At present, amplicon-sequencing studies-known also as metabarcoding studies, in which the primary data consist of targeted, amplified fragments of DNA sequenced from many taxa in a mixture-struggle to link genetic observations to the underlying biology in a quantitative way, but many applications require quantitative information about the taxa or systems under scrutiny. As metabarcoding studies proliferate in ecology, it becomes more important to develop ways to make them quantitative to ensure that their conclusions are adequately supported. Here we link previously disparate sets of techniques for making such data quantitative, showing that the underlying polymerase chain reaction mechanism explains the observed patterns of amplicon data in a general way. By modeling the process through which amplicon-sequence data arise, rather than transforming the data post hoc, we show how to estimate the starting DNA proportions from a mixture of many taxa. We illustrate how to calibrate the model using mock communities and apply the approach to simulated data and a series of empirical examples. Our approach opens the door to improve the use of metabarcoding data in a wide range of applications in ecology, public health, and related fields.

Metabarcoding dietary analysis in the insectivorous bat Nyctalusleisleri and implications for conservation.

In this study, we aim to uncover diet preferences for the insectivorous bat Nyctalusleisleri (Leisler's bat, the lesser noctule) and to provide recommendations for conservation of the species, based on the analysis of prey source habitats. Using a novel guano trap, we sampled bat faeces at selected roosts in a forest in Germany and tested two mitochondrial markers (COI and 16S) and three primer pairs for the metabarcoding of bat faecal pellets. We found a total of 17 arthropod prey orders comprising 358 species in N.leisleri guano. The most diverse orders were Lepidoptera (126 species), Diptera (86 species) and Coleoptera (48 species), followed by Hemiptera (28 species), Trichoptera (16 species), Neuroptera (15 species) and Ephemeroptera (10 species), with Lepidoptera species dominating in spring and Diptera in summer. Based on the ecological requirements of the most abundant arthropod species found in the bat guano, we propose some recommendations for the conservation of N.leisleri that are relevant for other insectivorous bat species.

Insights into the prey of Vespa mandarinia (Hymenoptera: Vespidae) in Washington state, obtained from metabarcoding of larval feces.

The northern giant hornet, Vespa mandarinia (Hymenoptera: Vespidae), was detected for the first time in North America in 2019. Four nests have since been located and removed in northwestern Washington State as part of an extensive survey and eradication program. This recent introduction into North America has prompted new research on the biology and ecology of V. mandarinia to help inform management strategies. In its native range, V. mandarinia is known to prey on a variety of insects including the economically important honey bee species Apis cerana and Apis mellifera. Although A. cerana has developed defense mechanisms against attack by V. mandarinia, A. mellifera have no such defenses and an entire hive can be quickly destroyed by only a few hornets. In North America the hornet has been observed foraging on paper wasps (Polistes dominula) and honey bees, but little else is known about prey use in its novel range. To address this knowledge gap, we employed a DNA metabarcoding approach to characterize species detected in larval feces collected from 3 of the 4 Washington V. mandarinia nests found to date. Sequences were recovered for 56 species across fourteen orders, of which 36 species were likely prey items and 20 were suspected inquilines. The most frequently detected species were other social Hymenoptera, with Dolichovespula maculata, P. dominula, and A. mellifera present in most samples. All of the species detected, except for A. mellifera, represent new prey records for V. mandarinia, with eight families of insects newly associated with giant hornets. These results suggest that V. mandarinia in Washington preys on an assortment of insects similar to those documented in its native range, and that this new invader has readily incorporated novel species into its foraging and diet.

Dietary and temporal partitioning facilitates coexistence of sympatric carnivores in the Everest region.

Carnivores, especially top predators, are important because they maintain the structure and function of ecosystems by top-down control. Exploring the coexistence between carnivores belonging to different ecological guilds can provide the data needed for the development of effective conservation strategies of endangered species. We used scats and camera traps to molecularly analyze the dietary composition of four predators that inhabit the Everest region and assess their activity patterns. Dietary analysis revealed 22 food Molecular Operational Taxonomic Units (MOTUs) of 7 orders and 2 classes. Snow leopard (Panthera uncia) and wolf (Canis lupus) had high dietary overlap (Pianka's index = 0.95), as they both mainly preyed on ungulates (%PR = 61%, 50%), while lynx (Lynx lynx) and red fox (Vulpes vulpes) mainly consumed small mammals (%PR = 62%, 76%). We observed lower dietary overlaps (Pianka's index = 0.53-0.70) between predators with large body size difference (snow leopard versus lynx, snow leopard versus red fox, wolf versus lynx, wolf versus fox), and dietary difference was significant (p < .01), proving dietary partitioning. In activity pattern analysis, predators exhibited higher temporal overlaps with the more frequently consumed prey species, showing that predator activity can be regulated by prey availability. We observed no obvious temporal avoidance between snow leopard and wolf because they had high activity overlap (Δ = 0.87). Red fox had the lowest coefficients of activity overlap with snow leopard and wolf (Δ = 0.60, 0.59), suggesting that fox tends to avoid snow leopard and wolf temporally. In this study, we revealed how dietary and temporal partitioning facilitates the coexistence of carnivores in Everest. These results will help to increase the understanding of coexistence mechanism of carnivore communities, and provide the scientific foundation for the conservation of wildlife living in the Qinghai-Tibet Plateau.

Investigating the link between morphological characteristics and diet in an island population of omnivorous reptiles (Sphenodon punctatus).

The morphological characteristics that impact feeding ecology in ectotherms, particularly reptiles, are poorly understood. We used morphometric measures and stable isotope analysis (carbon-13 and nitrogen-15) to assess the link between diet and functional morphology in an island population of an evolutionarily unique reptile, the tuatara (Sphenodon punctatus). First, we established a significant positive correlation between overall body size, gape size, and fat store in tuatara (n=56). Next, we describe the relationship between stable isotope profiles created from whole blood and nail trim samples and demonstrate that nail trims offer a low-impact method of creating a long-term dietary profile in ectotherms. We used nitrogen-15 values to assess trophic level in the population and found that tuatara on Takapourewa forage across multiple trophic levels. Finally, we found a significant relationship between gape size and carbon-13 (linear regression: P<0.001), with tuatara with large gapes showing dietary profiles that suggest a higher intake of marine (seabird) prey. However, whether body size or gape size is the primary adaptive characteristic allowing for more optimal foraging is yet unknown. This article has an associated First Person interview with the first author of the paper.

Dietary characterization of the endangered salt marsh harvest mouse and sympatric rodents using DNA metabarcoding.

The salt marsh harvest mouse (Reithrodontomys raviventris; RERA) is an endangered species endemic to the coastal wetlands of the San Francisco Estuary, California. RERA are specialized to saline coastal wetlands, and their historical range has been severely impacted by landscape conversion and the introduction of non-native plant and rodent species. A better understanding of their diet is needed to assess habitat quality, particularly in relation to potential competitors. We investigated three questions using DNA metabarcoding with ITS2 and trnL markers: (1) Do RERA specialize on the native plant, pickleweed (Salicornia pacifica), (2) Do RERA consume non-native plants, and (3) What is the dietary niche breadth and overlap with three sympatric native and non-native rodents? RERA diet was dominated by two plants, native Salicornia and non-native salt bush (Atriplex spp.), but included 48 plant genera. RERA diet breadth was narrowest in fall, when they consumed the highest frequencies of Salicornia and Atriplex, and broadest in spring, when the frequencies of these two plants were lowest. Diet breadth was slightly lower for RERA than for co-occurring species in pairwise comparisons. All four species consumed similarly high frequencies of wetland plants, but RERA consumed fewer grasses and upland plants, suggesting that it may be less suited to fragmented habitat than sympatric rodents. Diet overlap was lowest between RERA and the native California vole (Microtis californicus). In contrast, RERA diet overlapped substantially with the native western harvest mouse (R. megalotis) and non-native house mouse (Mus musculus), suggesting potential for competition if these species become sufficiently abundant.