Coral reef mesopredators switch prey,shortening food chains,in response to habitat degradation

Diet specificity is likely to be the key predictor of a predator's vulnerability to changing habitat and prey conditions. Understanding the degree to which predatory coral reef fishes adjust or maintain prey choice, in response to declines in coral cover and changes in prey availability, is critical for predicting how they may respond to reef habitat degradation. Here, we use stable isotope analyses to characterize the trophic structure of predator–prey interactions on coral reefs of the Keppel Island Group on the southern Great Barrier Reef, Australia. These reefs, previously typified by exceptionally high coral cover, have recently lost much of their coral cover due to coral bleaching and frequent inundation by sediment‐laden, freshwater flood plumes associated with increased rainfall patterns. Long‐term monitoring of these reefs demonstrates that, as coral cover declined, there has been a decrease in prey biomass, and a shift in dominant prey species from pelagic plankton‐feeding damselfishes to territorial benthic algal‐feeding damselfishes, resulting in differences in the principal carbon pathways in the food web. Using isotopes, we tested whether this changing prey availability could be detected in the diet of a mesopredator (coral grouper, Plectropomus maculatus). The δ¹³C signature in grouper tissue in the Keppel Islands shifted from a more pelagic to a more benthic signal, demonstrating a change in carbon sources aligning with the change in prey availability due to habitat degradation. Grouper with a more benthic carbon signature were also feeding at a lower trophic level, indicating a shortening in food chains. Further, we found a decline in the coral grouper population accompanying a decrease in total available prey biomass. Thus, while the ability to adapt diets could ameliorate the short‐term impacts of habitat degradation on mesopredators, long‐term effects may negatively impact mesopredator populations and alter the trophic structure of coral reef food webs.     

Trophic behaviour of juvenile reef fishes inhabiting interlinked mangrove–seagrass habitats in offshore mangrove islets

Stable isotope (δ¹³C and δ¹⁵N) and gut content analyses were used to investigate size‐related feeding habits of four reef fishes (the beaugregory Stegastes leucostictus, the french grunt Haemulon flavolineatum, the schoolmaster snapper Lutjanus apodus and the yellowtail snapper Ocyurus chrysurus) inhabiting an offshore (non‐estuarine) mangrove islet off Belize, Central America. Comparisons of isotopic niche space and Schoener diet similarity index suggested a low to moderate degree of niche overlap between fish size groups. The δ¹³C gradient between mangrove and seagrass prey as well as results of Bayesian mixing models revealed that sampled fishes relied mostly on seagrass prey items. Only small and large juveniles of the carnivorous species L. apodus derived a part of their diet from mangroves by targeting mangrove‐associated Grapsidae crabs and fish prey, respectively. Isotopic niche shifts were particularly obvious for carnivorous fishes that ingested larger prey items (Xanthidae crabs and fishes) during their ontogeny. The utilization of mangrove food resources is less than expected and depends on the ecology and life history of the fish species considered. This research highlights that mangrove‐derived carbon contributed relatively little to the diets of four fish taxa from an offshore mangrove islet.     

Food partitioning and spatial subsidy in shelter‐limited fishes inhabiting patchy reefs of Patagonia

The diets of the most conspicuous reef‐fish species from northern Patagonia, the carnivorous species Pseudopercis semifasciata, Acanthistius patachonicus, Pinguipes brasilianus and Sebastes oculatus were studied. Pinguipes brasilianus had the narrowest diet and most specialized feeding strategy, preying mostly on reef‐dwelling organisms such as sea urchins, limpets, bivalves, crabs and polychaetes. The diet of A. patachonicus was characterized by the presence of reef and soft‐bottom benthic organisms, mainly polychaetes, crabs and fishes. Pseudopercis semifasciata showed the broadest spectrum of prey items, preying upon reef, soft‐bottom and transient organism (mainly fishes, cephalopods and crabs). All S. oculatus guts were empty, but stable‐isotope analyses suggested that this species consumed small fishes and crabs. In general, P. brasilianus depended on local prey populations and ate different reef‐dwelling prey than the other species. Pseudopercis semifasciata, A. patachonicus and probably S. oculatus, however, had overlapping trophic niches and consumed resources from adjacent environments. The latter probably reduces the importance of food as a limiting resource for these reef‐fish populations, facilitating their coexistence in spite of their high trophic overlap.     

Sensory cues of a top‐predator indirectly control a reef fish mesopredator

Behavioural trophic cascades highlight the importance of indirect/risk effects in the maintenance of healthy trophic‐level links in complex ecosystems. However, there is limited understanding on how the loss of indirect top–down control can cascade through the food‐web to modify lower level predator–prey interactions. Using a reef fish food‐web, our study examines behavioural interactions among predators to assess how fear elicited by top‐predator cues (visual and chemical stimuli) can alter mesopredator behaviour and modify their interaction with resource prey. Under experimental conditions, the presence of any cue (visual, chemical, or both) from the top‐predator (coral trout Plectropomus leopardus) strongly restricted the distance swum, area explored and foraging activity of the mesopredator (dottyback Pseudochromis fuscus), while indirectly triggering a behavioural release of the resource prey (recruits of the damselfish Pomacentrus chrysurus). Interestingly, the presence of a large non‐predator species (thicklip wrasse Hemigymnus melapterus) also mediated the impact of the mesopredator on prey, as it provoked mesopredators to engage in an ‘inspection’ behaviour, while significantly reducing their feeding activity. Our study describes for the first time a three‐level behavioural cascade of coral reef fish and stresses the importance of indirect interactions in marine food‐webs.     

Combined gut‐content and stable isotope trophic analysis of the pelagic stingray Pteroplaytrygon violacea (Bonaparte, 1832) diet from the western North Atlantic Ocean

The understanding of trophic relationships is vital for correctly modeling ecosystems and ecosystem effects of fisheries removals. The pelagic stingray is found in epipelagic sub‐tropical and tropical waters worldwide and is a common bycatch in pelagic longline fisheries. Between August 2008 and November 2011, 156 specimens (81 males; 75 females) were collected during pelagic longline fishing operations in the US South Atlantic Bight and Gulf of Mexico. Stomach content analyses found that the major prey items were cephalopod molluscs (59.18%), followed by actinopterygiian fishes (37.75%), and decapod crustaceans (35.71%). These concentrations of prey items found in the stomachs coincide with previous studies done in the Pacific Ocean. In contrast to previous studies that found high percentages of empty stomachs (63%), the current percentage of empty stomachs was much lower (25.6%), likely due to shorter times between collection and inspection. Stable isotope analysis (δ¹³C and δ¹⁵N) was performed on white muscle in order to correlate the trophic position with gut‐content analysis. The δ¹³C values ranged from ‐18.81‰ to ‐16.70‰, while the δ¹⁵N ranged from 6.11‰ to 11.88‰. Modeling of stable isotope data suggest that while squid are occasionally an important part of the pelagic stingray diet, prey usually consist of shrimp and other pelagic crustaceans. Pelagic stingrays fed within two trophic levels, but their prey appeared to feed on different carbon sources than those found in other pelagic elasmobranchs. A deeper understanding of the pelagic stingray diet sources can help fisheries management as it begins to transition into ecosystem‐based management.     

Trophic ecology of three stingrays (Myliobatoidei: Dasyatidae) off the Brazilian north-eastern coast: Habitat use and resource partitioning

Understanding the ecological role of species with overlapping distributions is central to inform ecosystem management. Here we describe the diet, trophic level and habitat use of three sympatric stingrays, Hypanus guttatus, H. marianae and H. berthalutzae, through combined stomach content and stable isotope (δ¹³C and δ¹⁵N) analyses. Our integrated approach revealed that H. guttatus is a mesopredator that feeds on a diverse diet of benthic and epibenthic marine and estuarine organisms, principally bivalve molluscs, Alpheus shrimp and teleost fishes. Isotopic data supported movement of this species between marine and estuarine environments. H. berthalutzae is also a marine generalist feeder, but feeds primarily on teleost fishes and cephalopods, and consequently occupies a higher trophic level. In contrast, H. marianae is a mesopredator specialized on shrimps and polychaetas occurring only in the marine environment and occupying a low niche breadth. While niche overlap occurred, the three stingrays utilized the same prey resources at different rates and occupied distinct trophic niches, potentially limiting competition for resources and promoting coexistence. These combined data demonstrate that these three mesopredators perform different ecological roles in the ecosystems they occupy, limiting functional redundancy.     

Does a top predator reduce the predatory impact of an invasive mesopredator on an endangered rodent?

The mesopredator release hypothesis (MRH) predicts that reduced abundance of top‐order predators results in an increase in the abundance of smaller predators (mesopredators) due to a reduction in intra‐guild predation and competition. The irruption of mesopredators that follows the removal of top‐order predators can have detrimental impacts on the prey of the mesopredators. Here we investigated the mechanisms via which the presence of a top‐order predator can benefit prey species. We tested predictions made according to the MRH and foraging theory by contrasting the abundances of an invasive mesopredator (red fox Vulpes vulpes) and an endangered prey species (dusky hopping mouse Notomys fuscus), predator diets, and N. fuscus foraging behaviour in the presence and absence of a top‐predator (dingo Canis lupus dingo). As predicted by the MRH, foxes were more abundant where dingoes were absent. Dietary overlap between sympatric dingoes and foxes was extensive, and fox was recorded in 1 dingo scat possibly indicating intra‐guild predation. Notomys fuscus were more likely to occur in fox scats than dingo scats and as predicted by the MRH N. fuscus were less abundant in the absence of dingoes. The population increase of N. fuscus following rainfall was dampened in the absence of dingoes suggesting that mesopredator release can attenuate bottom‐up effects, although it remains conceivable that differences in grazing regimes associated with dingo exclusion could have also influenced N. fuscus abundance. Notomys fuscus exhibited lower giving‐up densities in the presence of dingoes, consistent with the prediction that their perceived risk of predation would be lower and foraging efficiency greater in the presence of a top‐predator. Our results suggest that mesopredator suppression by a top predator can create a safer environment for prey species where the frequency of fatal encounters between predators and prey is reduced and the non‐consumptive effects of predators are lower.     

Energy flow to two abundant consumers in a subtropical oyster reef food web

Oyster reefs are among the most threatened coastal habitat types, but still provide critical habitat and food resources for many estuarine species. The structure of oyster reef food webs is an important framework from which to examine the role of these reefs in supporting high densities of associated fishes. We identified major trophic pathways to two abundant consumers, gray snapper (Lutjanus griseus) and crested goby (Lophogobius cyprinoides), from a subtropical oyster reef using stomach content and stable isotope analysis. The diet of gray snapper was dominated by crabs, with shrimp and fishes also important. Juvenile gray snapper fed almost entirely on oyster reef-associated prey items, while subadults fed on both oyster reef- and mangrove-associated prey. Based on trophic guilds of the gray snapper prey, as well as relative δ¹³C values, microphytobenthos is the most likely basal resource pool supporting gray snapper production on oyster reefs. Crested goby had omnivorous diets dominated by bivalves, small crabs, detritus, and algae, and thus were able to take advantage of prey relying on production from sestonic, as well as microphytobenthos, source pools. In this way, crested goby represent a critical link of sestonic production to higher trophic levels. These results highlight major trophic pathways supporting secondary production in oyster reef habitat, thereby elucidating the feeding relationships that render oyster reef critical habitat for many ecologically and economically important fish species.     

Utility of fish scales from stock assessment surveys in stable isotope analysis for initial assessments of trophic relationships in riverine fish communities

The utility of using fish scales collected during stock assessment exercises to assess the trophic relationships of riverine fishes using their stable isotopes of d¹³C and d¹⁵N was tested using three riverine fish communities in England (Rivers Great Ouse, Ivel and Goyt). In each river, European barbel Barbus barbus was an important species, with other cyprinid species, including chub Squalius cephalus, present. Stable isotope analyses was completed using relatively small sample sizes per species (<11) from fish samples collected in 2001, 2005 and 2006 when up to 5 scales were collected from each fish. The calculation of standard ellipse areas (as a measure of trophic niche size) revealed that relative to other fishes, B. barbus occupied high trophic positions with minimal overlap in their trophic niche with other species, especially S. cephalus. As the analysed fish samples comprised species of various length ranges and as length has strong ontogenetic consequences for fish diet composition, generalized linear models were developed in which length was the covariate; model outputs included length‐adjusted mean δ¹³C and δ¹⁵N for each species. In each fish community, significant differences in δ¹³C and δ¹⁵N were apparent between B. barbus and S. cephalus, but were less apparent between B. barbus and other fishes. Thus, whilst the utility of using fish scales from stock assessments in stable isotope analyses are limited due to the differing length ranges of the sampled fishes, they can be useful in identifying trophic differences between species when methods such as stomach content analyses are unavailable.     

Trophic relationships between the larvae of two freshwater mussels and their fish hosts

Freshwater mussels of the order Unionoida have life cycles that include larval attachment to and later metamorphosis on suitable host fishes. Information on the trophic relationship between unionoid larvae and their host fishes is scarce. We investigated the trophic interaction between fish hosts and encysted larvae of two species of freshwater mussels, Margaritifera margaritifera and Unio crassus, using stable isotope analyses of larvae and juvenile mussels as well as of host fish gill and muscle tissues before and after infestation. Due to different life histories and durations of host‐encystment, mass and size increase in M. margaritifera during the host‐dependent phase were greater than those of U. crassus. δ¹³C and δ¹⁵N signatures of juvenile mussels approached isotopic signatures of fish tissues, indicating a parasitic relationship between mussels and their hosts. Shifts were more pronounced for M. margaritifera, which had a five‐fold longer host‐dependent phase than U. crassus. The results of this study suggest that stable isotope analyses are a valuable tool for characterizing trophic relationships and life history strategies in host–parasite systems. In the case of unionoid mussels, stable isotopic shifts of the larvae are indicative of the nutritional versus phoretic importance of the host.     

Stable isotope analysis indicates trophic differences among forest floor carabids in Japan

Differences in trophic niches among carabid beetles (Coleoptera: Carabidae) co‐occurring on the forest floors of warm temperate forests in central Japan were studied using carbon (δ¹³C) and nitrogen (δ¹⁵N) stable isotope analyses. Different carabid species showed similar δ¹⁵N values, which were higher than those of their possible invertebrate prey (herbivores and detritivores) collected from the litter layer, indicating that these species were consumers in the same trophic level. In contrast, δ¹³C values differed among carabid species, indicating interspecific differences in prey animals. The variation in the δ¹³C value was larger in summer than in autumn. In summer, δ¹³C values indicated that some carabids depended highly on either grazing (low δ¹³C values) or detrital sources (high δ¹³C values) within the food chain [Chlaenius posticalis Motschulsky and Haplochlaenius costiger (Chaudoir), respectively], although other species with intermediate δ¹³C values likely depended on both. The latter group of species comprised mostly two dominant genera (Carabus and Synuchus). Although congeners might have similar feeding habits, the stable isotope ratios indicated trophic niche differences between adults of different species and between adults and larvae of the same genus.     

Predicting the contributions of novel marine prey resources from angling and anadromy to the diet of a freshwater apex predator

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Estimating the role of three mesopredatory fishes in coral reef food webs at Ningaloo Reef,Western Australia

Within the complex food webs that occur on coral reefs, mesopredatory fish consume small-bodied prey and transfer accumulated biomass to other trophic levels. We estimated biomass, growth and mortality rates of three common mesopredators from Ningaloo Reef in Western Australia to calculate their annual turnover rates and potential contribution to local trophic dynamics. Biomass estimates of the serranid Epinephelus rivulatus (4.46 ± 0.76 g m⁻²) were an order of magnitude greater than two smaller-bodied mesopredatory fishes, Pseudochromis fuscus (0.10 ± 0.03 g m⁻²) and Parapercis clathrata (0.23 ± 0.31 g m⁻²). Growth parameters generated from a von Bertalanffy growth function fitted to size-at-age data, however, indicated that mortality rates for the three mesopredators were similar and that 32–55 % of fish survived each year. Consequently, interspecific differences in annual turnover rates among E. rivulatus (1.9 g m⁻² yr⁻¹), Pa. clathrata (0.10 g m⁻² yr⁻¹) and Ps. fuscus (0.07 g m⁻² yr⁻¹) were an artefact of differences in local biomass estimates. The rapid turnover estimates for E. rivulatus suggest this species is an important conduit of energy within the isolated patch reef habitat where it is typically found, while Ps. fuscus and Pa. clathrata channel smaller amounts of energy from specific habitats in the Ningaloo lagoon. Apparent differences in habitat, diet and turnover rates of the three species examined provide an insight into the different roles these species play in coral reef food webs and suggest that life-history traits allow for variability in the local and spatial contribution of these species at Ningaloo Reef. Moreover, calculating turnover rates of a broader suite of fish species from a range of trophic groups will help better define the role of fishes in coral reef trophic dynamics.

     

Feeding strategy of silverside species in eutrophic Lake Pátzcuaro,Mexico

The trophic guild, niche breadth, and trophic position of two different ontogenetic phases were measured according to the size of Chirostoma spp. captured in different zones of Lake Pátzcuaro, Mexico. Analyses were carried out at the genus level due the difficulties to find diagnostic characters to separate the three small silverside species accurately, since even the molecular analyses of mitochondrial DNA for a subsample of the most morphologically differentiated specimens were conducted without success. Fish were captured with a seine net during the wet (September and November 2009) and dry (February and June 2010) seasons at four environmentally different sites. Gut contents of 480 Chirostoma specimens were divided into size‐class intervals of 38–51 mm and 66–71 mm for analyses. The contribution of each food item was quantified using the frequency of occurrence and area percentage. Fish were categorized by size, and diets were compared between fish sizes and sites. Dorsal muscle tissue and zooplankton were obtained for nitrogen isotope signature (δ¹⁵N) analyses. The trophic guild was determined using a modified quadrant method for gut content analysis; the importance of each prey item was estimated according to a modified index of relative importance. Niche breadth was calculated by the Levins’ Index. The trophic position was assessed using the TrophLab Program and stable isotope analysis. The index of relative importance among study sites and fish size classes was determined by a multivariate approach. Silversides represent a secondary and tertiary consumer in Lake Pátzcuaro that feed mainly on evasive prey, such as copepods (55%) and on non‐evasive prey, such as cladocerans (25%) at sites with the lowest transparency in the water column (from 350 mm to 150 mm). Fish scales (11%) and insects (6%) were secondary food items. Ingestion of juvenile silversides was rare (1%). Chirostoma spp. prey on insects on the lake bottom and on water hyacinth roots. Based on δ¹⁵N analysis, the trophic position of silversides changes between wet and dry seasons. Their opportunistic and specialized predatory feeding strategy indicates the capability of silversides to resist anthropic changes in the lake.     

Comparative feeding ecology of invasive Ponto-Caspian gobies

Invasions of Ponto-Caspian gobiid fishes are suspected to cause regime shifts in freshwater ecosystems. This study compared the trophic niche differentiations of Neogobius melanostomus and Ponticola kessleri in the upper Danube River using stable isotope analyses (δ¹³C and δ¹⁵N), gut content analyses and morphometric analyses of the digestive tract. Both species were identified as predacious omnivores with high dietary overlap and a generalistic feeding strategy. Amphipods (especially invasive Dikerogammarus spp.) contributed 2/3 to the index of food importance. δ¹⁵N-signatures of N. melanostomus revealed an ontogenetic diet shift and significantly exceeded those in P. kessleri by ~1.5‰, indicating a niche separation of half a trophic level. P. kessleri had shorter uncoiled intestinal tracts than N. melanostomus, indicating a narrower niche and adaptation to animal food. Trophic niches in both species expanded during the growth period with increasing intraguild predation and cannibalism in P. kessleri and increasing molluscivory in N. melanostomus. P. kessleri showed a higher degree of specialization and more stable feeding patterns across seasons, whereas N. melanostomus adapted its diet according to the natural prey availability. The feeding patterns of both species observed in the upper Danube River strongly differ from those in their native ranges, underlining their great plasticity. Both goby species consumed mainly other non-native species (~92% of gut contents) and seemed to benefit from previous invasions of prey species like Dikerogammarus villosus. The invasive success of gobies and their prey mirror fundamental ecological changes in large European freshwater ecosystems.     

Prey composition and ontogenetic shift in coastal populations of longnose gar Lepisosteus osseus

Longnose gar Lepisosteus osseus were collected from May 2012 to July 2013 in the Charleston Harbor and Winyah Bay estuaries (SC, U.S.A.). This study examined trends in stomach fullness, described major prey components and their importance in the diet of L. osseus, compared stomach content‐based trophic level estimates with the stable‐isotope‐based proxy: δ¹⁵N and tested for the occurrence of an ontogenetic diet shift using stomach content analysis and stable C and N isotopes (δ¹³C and δ¹⁵N). Dominant prey families were Clupeidae, Sciaenidae, Penaeidae, Fundulidae and Mugilidae, with the highest consumption rates in autumn. Trophic levels calculated using stomach contents did not correspond to δ¹⁵N (P > 0·05). Stomach contents and stable‐isotope signatures indicate ontogenetic prey composition shifts from low trophic level benthic prey (fundulids) to higher trophic level pelagic prey (clupeids) as the fish grow between 400 and 600 mm in standard length. Due to their biomass, abundance and top predator status, L. osseus play a significant ecological role in the estuarine community composition, although this effect has often been overlooked by past researchers and should be considered in future estuarine community studies.     

Trophic shift in young‐of‐the‐year Mugilidae during salt‐marsh colonization

This study investigated the trophic shift of young‐of‐the‐year (YOY) thinlip grey mullet Liza ramada and golden grey mullet Liza aurata during their recruitment in a salt marsh located on the European Atlantic Ocean coast. Stable‐isotope signatures (δ¹³C and δ¹⁵N) of the fishes followed a pattern, having enrichments in ¹³C and ¹⁵N with increasing fork length (L_F): δ¹³C in fishes < 30 mm ranged from ?19.5 to ?15.0‰, whereas in fishes > 30 mm δ¹³C ranged from ?15.8 to ?12.7‰, closer to the level in salt‐marsh food resources. Large differences between the δ¹⁵N values of mugilids and those of food sources (6·0‰ on average) showed that YOY are secondary consumers, similar to older individuals, when feeding in the salt marsh. YOY mugilids shift from browsing on pelagic prey to grazing on benthic resources from the salt marsh before reaching 30 mm L_F. The results highlight the role of European salt marshes as nurseries for juvenile mugilids.     

Trophic interactions in an ant nest microcosm: a combined experimental and stable isotope (δ13C/δ15N) approach

Living in close association with other organisms has proven to be a widespread and successful strategy in nature. Some communities are completely driven by symbiotic associations and therefore, intimate relationships among the partners can be expected. Here, we analyzed in‐depth the food web of a particularly rich community of arthropods found in strict association with European red wood ants (Formica rufa group). We studied the trophic links between different ant‐associated myrmecophiles and food sources associated with the host ant, but also tested predator–prey links among myrmecophiles themselves. Our approach combined direct feeding tests and stable carbon and nitrogen isotope analyses for a large number of myrmecophiles. The results of the direct feeding tests reveal a complex food web. Most myrmecophiles were found to parasitize on ant brood. Moreover, we encountered multiple trophic predator– prey links among the myrmecophiles. The results of the stable isotope analyses complement these findings and indicate the existence of multiple trophic levels and trophic isotopic niche compartmentalization. δ¹⁵N values were strongly correlated with the trophic levels based on the direct tests, reflecting that δ¹⁵N values of myrmecophiles increased with higher trophic levels. This strong correlation underlines the strength of stable isotopes as a powerful tool to assess trophic levels. In addition, the stable isotope data suggest that most species only facultatively prey on ant brood. The presence of numerous trophic interactions among symbionts clearly contrasts with the traditional view of social insects nests as offering an enemy‐free space for symbionts. Interestingly, the ant host can indirectly benefit from these interactions because brood predators are also preyed upon by other myrmecophiles. Overall, this study provides unique insights into the complex interactions in a small symbiont microcosm system and suggests that the interactions between host and symbiont might be mediated by other symbionts in the same community.     

Comparing pellet and stable isotope analyses of nestling Bonelli's Eagle Aquila fasciata diet

Diet analyses are central to the study of avian trophic ecology, and stable isotope analyses have made an increasing contribution in the last two decades. Few isotopic studies have assessed the diet of raptor species, which are more frequently analysed by conventional diet methods such as pellet analysis. In this study, we compare prey consumption estimates of nestling Bonelli's Eagles Aquila fasciata from conventional pellet analysis (in terms of items and biomass) and stable isotopic mixing models (SIAR) using δ¹³C, δ¹⁵N and δ³⁴S of feathers. The pellet analysis showed that European Rabbits Oryctolagus cuniculus, pigeons (mainly Common Wood Pigeons Columba palumbus and Domestic Pigeons Columba livia dom.), Red‐legged Partridges Alectoris rufa, passerines, Yellow‐legged Gulls Larus michahellis and Eurasian Red Squirrels Sciurus vulgaris were the main prey, so they were selected for diet reconstructions in SIAR. At the population level, mean prey consumption estimates were similar for pellets (both items and biomass) and SIAR. At the territory level, the weighted kappa statistic showed good ordinal scale agreement in main prey consumption between items or biomass and SIAR. Although the intraclass correlation coefficient showed poor method agreement when considering all prey in the same analysis, the intraclass correlation coefficients for each prey category showed significant agreement between pellets and SIAR when estimating the consumption of Rabbits, pigeons and Gulls, with lower agreement for passerines and Squirrels. Lastly, there was poor method agreement for estimates of Partridges. Our results suggest an overall agreement between the pellet analysis and SIAR when estimating nestling Bonelli's Eagle diet at both the population and, to a lesser extent, the territory level, supporting the usefulness of isotopic mixing models when identifying the terrestrial and marine components of raptor diets.     

Refining trophic dynamics through multi‐factor Bayesian mixing models: A case study of subterranean beetles

Food web dynamics are vital in shaping the functional ecology of ecosystems. However, trophic ecology is still in its infancy in groundwater ecosystems due to the cryptic nature of these environments. To unravel trophic interactions between subterranean biota, we applied an interdisciplinary Bayesian mixing model design (multi‐factor BMM) based on the integration of faunal C and N bulk tissue stable isotope data (δ¹³C and δ¹⁵N) with radiocarbon data (Δ¹⁴C), and prior information from metagenomic analyses. We further compared outcomes from multi‐factor BMM with a conventional isotope double proxy mixing model (SIA BMM), triple proxy (δ¹³C, δ¹⁵N, and Δ¹⁴C, multi‐proxy BMM), and double proxy combined with DNA prior information (SIA + DNA BMM) designs. Three species of subterranean beetles (Paroster macrosturtensis, Paroster mesosturtensis, and Paroster microsturtensis) and their main prey items Chiltoniidae amphipods (AM1: Scutachiltonia axfordi and AM2: Yilgarniella sturtensis), cyclopoids and harpacticoids from a calcrete in Western Australia were targeted. Diet estimations from stable isotope only models (SIA BMM) indicated homogeneous patterns with modest preferences for amphipods as prey items. Multi‐proxy BMM suggested increased—and species‐specific—predatory pressures on amphipods coupled with high rates of scavenging/predation on sister species. SIA + DNA BMM showed marked preferences for amphipods AM1 and AM2, and reduced interspecific scavenging/predation on Paroster species. Multi‐factorial BMM revealed the most precise estimations (lower overall SD and very marginal beetles' interspecific interactions), indicating consistent preferences for amphipods AM1 in all the beetles' diets. Incorporation of genetic priors allowed crucial refining of the feeding preferences, while integration of more expensive radiocarbon data as a third proxy (when combined with genetic data) produced more precise outcomes but close dietary reconstruction to that from SIA + DNA BMM. Further multidisciplinary modeling from other groundwater environments will help elucidate the potential behind these designs and bring light to the feeding ecology of one the most vital ecosystems worldwide.