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Dietary analysis using DNA metabarcoding is a powerful tool that is increasingly being used to further our knowledge of trophic interactions in highly complex food webs but is not without limitations. Omnivores, the most generalist of consumers, pose unique challenges when using such methods. Here, we provide the rationale to understand the problems associated with analysing the complex diets of omnivores. By reviewing existing metabarcoding studies of omnivorous diet, and constructing hypothetical scenarios arising from each, we outline that great caution is required when interpreting sequencing data in such cases. In essence, the problems of accidental consumption and secondary ingestion are significant sources of error when investigating omnivorous diets. The integration of multiple high throughput sequencing markers increases the taxonomic breadth of taxa detected but we reveal how some detections may be misleading. Disentangling which taxa have been deliberately or accidentally consumed by the focal omnivore is challenging and can falsely emphasise those that were not intentionally consumed, obscuring biologically meaningful interactions. Although we suggest ways to disentangle these issues, we urge that the results of such analyses should be interpreted with caution and all possible scenarios for the presence of biota within omnivores given due consideration. [ABSTRACT FROM AUTHOR]

Non‐native ants can cause ecosystem‐wide ecological change, and these changes are generally assumed to be negative. Despite this, the evidence base has never been holistically synthesised to quantify whether and to what degree non‐native ants impact native species diversity.In this study, we performed a meta‐analysis of the effects of ant invasion on animal communities. We extracted data from 46 published articles investigating abundance (156 effect sizes) and richness (53 effect sizes) responses of animal taxa to ant invasion in locations relatively unimpacted by other stressors (e.g. human disturbance, other non‐native species) to help isolate the effects of invasion.Overall, local animal diversity declined severely, with species abundance and richness lower by 42.79% and 53.56%, respectively, in areas with non‐native ants compared with intact uninvaded sites. We then combined responses of individual animal taxa extracted from an article into a single response to represent the 'community' abundance (40 effect sizes) or richness (28 effect sizes) response to non‐native ants represented in each article. Local communities decreased substantially in total abundance (52.67%) and species richness (53.47%) in invaded sites.These results highlight non‐native ants as the drivers, rather than passengers, of large net‐negative reductions to animal community diversity in relatively undisturbed systems around the world, approximately halving local species abundance and richness in invaded areas. Improved international prevention processes, early detection systems harnessing emerging technologies, and well‐designed control measures deployable by conservation practitioners are urgently needed if these effects are to be mitigated, prevented or reversed. [ABSTRACT FROM AUTHOR]

Variation in prey resources influences the diet and behaviour of predators. When prey become limiting, predators may travel farther to find preferred food or adjust to existing local resources. When predators are habitat limited, local resource abundance impacts foraging success. We analysed the diet of Myotis lucifugus (little brown bats) from Nova Scotia (eastern Canada) to the Northwest Territories (north-western Canada). This distribution includes extremes of season length and temperature and encompasses colonies on rural monoculture farms, and in urban and unmodified areas. We recognized nearly 600 distinct species of prey, of which ≈30% could be identified using reference sequence libraries. We found a higher than expected use of lepidopterans, which comprised a range of dietary richness from ≈35% early in the summer to ≈55% by late summer. Diptera were the second largest prey group consumed, representing ≈45% of dietary diversity early in the summer. We observed extreme local dietary variability and variation among seasons and years. Based on the species of insects that were consumed, we observed that two locations support prey species with extremely low pollution and acidification tolerances, suggesting that these are areas without environmental contamination. We conclude that there is significant local population variability in little brown bat diet that is likely driven by seasonal and geographical changes in insect diversity, and that this prey may be a good indicator of environment quality. [ABSTRACT FROM AUTHOR]

An introduction to the journal is presented in which the authors discuss the molecular detection of trophic interactions between predators and their preys.

Prevalence and intensity of parasitic infections are often higher in male than in female vertebrates. This bias may represent either differences between host sex in exposure or susceptibility to parasites. The former may be due to sex-specific behaviour of the host, including differential habitat use or diet. Differences in susceptibility are often regarded as a negative effect of male sex steroid hormones on the immune system. Host-parasite dynamics are of great interest in terms of reptile survival, ecology and conservation. We used, for the first time, molecular diagnostics to track nematode parasitism in wild populations of reptiles noninvasively. Using slow worms ( Anguis fragilis) as a model species, we investigated the interacting effects of time of year, sex, length, weight and climatic variables on the prevalence of the gastroenterological parasitic nematode Neoxysomatium brevicaudatum. Faeces were collected from three sites over 2 years. There was an interaction between sex and time of year, with lower nematode prevalence in males than in females in July or August (different between years) but a high prevalence in males in April. As the latter is during the slow worm breeding season, this may be the result of testosterone-induced immunosuppression. A second-order interaction between slow worm length and weight was found to be significant, with a positive association between prevalence and body condition in young slow worms and a negative association in older slow worms. The convex pattern of nematode prevalence with age that emerged suggests an increase with age-related exposure and a decrease with age-related acquired immunity. [ABSTRACT FROM AUTHOR]

Generalist species with numerous food web interactions are thought to provide stability to ecosystem dynamics; however, it is not always clear whether habitat generality translates into dietary diversity. Big brown bats are common across North America and employ a flexible foraging strategy over water, dense forests, forest edges and rural and urban settings. Despite this generalist use of habitat, they are paradoxically characterized as beetle specialists. However, hard carapaces may preferentially survive digestion leading to over-representation during morphological analysis of diet. This specialization has not been evaluated independently using molecular analysis and species-level identification of prey. We used next-generation sequencing to assess the diet of big brown bats. Beetles were consumed in the highest frequency but Lepidoptera species richness was highest among identified prey. The consumption of species showed strong seasonal and annual variation. While Coleoptera consumption varied, Lepidoptera and Ephemeroptera were relatively constant dietary components. Dietary diversity increased in late summer when insect diversity decreases. Our results indicate that big brown bats are dietary generalists and, while beetles are an important component of the diet, Lepidoptera are equally important, and Lepidoptera and Ephemeroptera are the only stable prey resource exploited. As resources become limited, big brown bats may respond by increasing the species richness of prey and thus their connectedness in the ecosystem. This characterization of diet corresponds well with a generalist approach to foraging, making them an important species in encouraging and maintaining ecosystem stability. [ABSTRACT FROM AUTHOR]

Summary1. Reports are reviewed of gastropod feeding (malacophagy) by spiders and harvestmen. Although the standard textbooks on arachnids recognise the importance of gastropods as prey of harvestmen, none apparently refers to malacophagy by spiders. A review of several hundred papers on spider feeding habits revealed that species from several families kill and devour slugs and snails in the laboratory and/or field. 2. Malacophagy has been reported most frequently among mygalomorph spiders, and can make up a substantial proportion of the diets of some species, however gastropods make up an insignificant percentage of the prey of most araneomorph spiders. The spiders that eat gastropods are species with broad diets composed predominantly of arthropod prey. No species of spider appears to feed exclusively on gastropod prey. 3. Harvestmen from several families have broad diets that often include gastropods. Several species of the family Trogulidae and at least one species of the family Ischyropsalididae [Ischyropsalis hellwigi (Panzer)] are specialised gastropod predators. The trogulids are slender animals that attack the snail through the shell aperture (shell intruders). Ischyropsalis hellwigi, on the other hand, can crush snail shells with its powerful chelicerae (shell breakers). 4. The review highlights apparent convergent evolution by harvestmen and Carabidae of two mutually exclusive morphologies found among gastropod predators. It also suggests that there is an urgent need for systematic studies to be conducted to establish the extent and ecological importance of malacophagy in natural and anthropogenically altered habitats. [ABSTRACT FROM AUTHOR]

Null models provide a valuable baseline against which fundamental ecological hypotheses can be tested and foraging choices that cannot be explained by neutral processes or sampling biases can be highlighted. In this way, null models can advance our understanding beyond simplistic dietary descriptions to identify drivers of interactions. This method, however, requires estimates of resource availability, which are generally imperfect representations of highly dynamic systems. Optimising method selection is crucial for study design, but the precise effects of different resource availability data on the efficacy of null models are poorly understood.Using spider–prey networks as a model, we used prey abundance (suction sample) and activity density (sticky trap) data, and combinations of the two, to simulate null networks. We compared null diet composition, network properties (e.g., connectance and nestedness) and deviations of simulations from metabarcoding‐based spider dietary data to ascertain how different prey availability data alter ecological interpretation.Different sampling methods produced different null networks and inferred distinct prey selectivity. Null models based on prey abundance and combined frequency‐of‐occurrence data generated null diet compositions, which more closely resembled the diet composition determined by metabarcoding. Null models based on prey abundance, activity density and proportionally combined data generated null network properties most like the networks constructed via dietary metabarcoding.We show that survey method choice impacts all aspects of null network analyses, the precise effects varying between methods but ultimately altering ecological interpretation by increasing disparity in network properties or trophic niches between null and directly constructed networks. Merging datasets can generate more complete prey availability data but is not a panacea because it introduces different biases. The choice of method should reflect the research hypotheses and study system being investigated. Ultimately, survey methods should emulate the foraging mode of the focal predator as closely as possible, informed by the known ecology, natural history and behaviour of the predator. [ABSTRACT FROM AUTHOR]

DNA metabarcoding is a technique used to survey biodiversity in many ecological settings, but there are doubts about whether it can provide quantitative results, that is, the proportions of each species in the mixture as opposed to a species list. While there are several experimental studies that report quantitative metabarcoding results, there are a similar number that fail to do so. Here, we provide the rationale to understand under what circumstances the technique can be quantitative. In essence, we simulate a mixture of DNA of S species with a defined initial abundance distribution. In the simulated PCR, each species increases its concentration following a certain amplification efficiency. The final DNA concentration will reflect the initial one when the efficiency is similar for all species; otherwise, the initial and final DNA concentrations would be poorly related. Although there are many known factors that modulate amplification efficiency, we focused on the number of primer–template mismatches, arguably the most important one. We used 15 common primers pairs targeting the mitochondrial COI region and the mitogenomes of ca. 1,200 insect species. The results showed that some primers pairs produced quantitative results under most circumstances, whereas some other primers failed to do so. In conclusion, depending on the primer pair used in the PCR amplification and on the characteristics of the mixture analysed (i.e., high species richness, low evenness), DNA metabarcoding can provide a quantitative estimate of the relative abundances of different species. [ABSTRACT FROM AUTHOR]

Generalist invertebrate predators are sensitive to weather conditions, but the relationship between their trophic interactions and weather is poorly understood. This study investigates how weather affects the identity and frequency of spider trophic interactions over time, alongside prey community structure, web characteristics and prey choice. Spiders (Linyphiidae and Lycosidae) and their prey were collected from barley fields in Wales, UK, from April to September 2017-2018. The gut contents of 300 spiders were screened using DNA metabarcoding, analysed via multivariate models and compared against prey availability using null models. When linyphiids were collected from webs, the height and area of webs were recorded and compared against weather conditions. Trophic interactions changed over time and with weather conditions, primarily related to concomitant changes in prey communities. Spiders did, however, appear to mitigate the effects of structural changes in prey communities through changing prey preferences according to prevailing weather conditions, possibly facilitated by adaptive web construction. Using these findings, we demonstrate that prey choice data collected under different weather conditions can be used to refine inter-annual predictions of spider trophic interactions, although prey abundance was secondary to diversity in driving the diet of these spiders. By improving our understanding of the interaction between trophic interactions and weather, we can better predict how ecological networks are likely to change over time in response to variation in weather conditions and, more urgently, global climate change. [ABSTRACT FROM AUTHOR]

Avian diet can be affected by site‐specific variables, such as habitat, as well as intrinsic factors such as sex. This can lead to dietary niche separation, which reduces competition between individuals, as well as impacting how well avian species can adapt to environmental variation. Estimating dietary niche separation is challenging, due largely to difficulties in accurately identifying food taxa consumed. Consequently, there is limited knowledge of the diets of woodland bird species, many of which are undergoing serious population declines. Here, we show the effectiveness of multi‐marker fecal metabarcoding to provide in‐depth dietary analysis of a declining passerine in the UK, the Hawfinch (Coccothraustes coccothraustes). We collected fecal samples from (n = 262) UK Hawfinches prior to, and during, the breeding seasons in 2016–2019. We detected 49 and 90 plant and invertebrate taxa, respectively. We found Hawfinch diet varied spatially, as well as between sexes, indicating broad dietary plasticity and the ability of Hawfinches to utilize multiple resources within their foraging environments. [ABSTRACT FROM AUTHOR]

Eurasian otters are apex predators of freshwater ecosystems and a recovering species across much of their European range; investigating the dietary variation of this predator over time and space, therefore, provides opportunities to identify changes in freshwater trophic interactions and factors influencing the conservation of otter populations. Here we sampled feces from 300 dead otters across England and Wales between 2007 and 2016, conducting both morphological analyses of prey remains and dietary DNA metabarcoding. Comparison of these methods showed that greater taxonomic resolution and breadth could be achieved using DNA metabarcoding but combining data from both methodologies gave the most comprehensive dietary description. All otter demographics exploited a broad range of taxa and variation likely reflected changes in prey distributions and availability across the landscape. This study provides novel insights into the trophic generalism and adaptability of otters across Britain, which is likely to have aided their recent population recovery, and may increase their resilience to future environmental changes. [ABSTRACT FROM AUTHOR]

The use of ecological replacements (analogue species to replace extinct taxa) to restore ecosystem functioning is a promising conservation tool. However, this approach is controversial, in part due to a paucity of data on interactions between analogue species and established taxa in the ecosystem.We conducted ecological surveys, comprehensively DNA barcoded an ecosystem's flora and inferred the diet of the introduced Aldabra giant tortoise, acting as an ecological replacement, to understand how it might have modified island plant communities on a Mauritian islet. Through further dietary analyses, we investigated consequential effects on the threatened endemic Telfair's skink.Dietary overlap between tortoises and skinks was greater than expected by chance. However, there was a negative correlation between tortoise and skink preferences in herbivory and minimal overlap in the plants most frequently consumed by the reptiles.Changes in the plant community associated with 7 years of tortoise grazing were characterised by a decrease in the percentage cover of native herbs and creepers, and an increase in the cover of an invasive herb when compared to areas without tortoises. However, tortoise dietary preferences themselves did not directly drive changes in the plant community.Tortoises successfully dispersed the seeds of an endemic palm, which in time may increase the extent of unique palm‐rich habitat. We found no evidence that tortoises have increased the extent of plant species hypothesised to be part of a lost Mauritian tortoise grazed community.Synthesis and applications. Due to a negative correlation in tortoise and skink dietary preferences and minimal overlap in the most frequently consumed taxa, the presence of tortoises is unlikely to have detrimental impacts on Telfair's skinks. Tortoise presence is likely to be beneficial to skinks in the long term by increasing the extent of palm‐rich habitat. Although tortoises are likely to play a role in controlling invasive plants, they are not a panacea for this challenge. After 7 years, tortoises have not resurrected a lost tortoise grazed community that we hypothesise might have existed in limited areas on the islet, indicating that further interventions may be required to restore this plant community. [ABSTRACT FROM AUTHOR]

The diet of an individual animal is subject to change over time, both in response to short‐term food fluctuations and over longer time scales as an individual ages and meets different challenges over its life cycle. A metabarcoding approach was used to elucidate the diet of different life stages of a migratory songbird, the Eurasian reed warbler (Acrocephalus scirpaceus) over the 2017 summer breeding season in Somerset, the United Kingdom. The feces of adult, juvenile, and nestling warblers were screened for invertebrate DNA, enabling the identification of prey species. Dietary analysis was coupled with monitoring of Diptera in the field using yellow sticky traps. Seasonal changes in warbler diet were subtle, whereas age class had a greater influence on overall diet composition. Age classes showed high dietary overlap, but significant dietary differences were mediated through the selection of prey; (i) from different taxonomic groups, (ii) with different habitat origins (aquatic vs. terrestrial), and (iii) of different average approximate sizes. Our results highlight the value of metabarcoding data for enhancing ecological studies of insectivores in dynamic environments. [ABSTRACT FROM AUTHOR]

Reptiles are declining in many parts of the world, mainly due to habitat loss and environmental change. A major factor in this is availability of suitable food. For many animals, dietary requirements shift during developmental stages and a habitat will only be suitable for conserving a species if it supports all stages. Conventional methods for establishing diet often rely on visual recognition of morphologically identifiable features of prey in faeces, regurgitation or stomach contents, which suffer from biases and poor resolution of taxa. DNA-based techniques facilitate noninvasive analysis of diet from faeces without these constraints. We tested the hypothesis that diet changes during growth stages of smooth snakes ( Coronella austriaca), which have a highly restricted distribution in the UK but are widespread in continental Europe. Small numbers of the sympatric grass snake ( Natrix natrix) were analysed for comparison. Faecal samples were collected from snakes and prey DNA analysed using PCR, targeting amphibians, reptiles, mammals and invertebrates. Over 85% of smooth snakes were found to have eaten reptiles and 28% had eaten mammals. Predation on mammals increased with age and was entirely absent among juveniles and subadults. Predation on reptiles did not change ontogenetically. Smooth snakes may, therefore, be restricted to areas of sufficiently high reptile densities to support young snakes. [ABSTRACT FROM AUTHOR]

In this investigation, we use variation in wing morphology, echolocation behaviour, patterns of habitat use and molecular diet analysis to demonstrate that six species of sympatric insectivorous bats in Jamaica show significant differences that could explain resource partitioning among the species. High-intensity echolocating species that used shorter, broadband signals and had shorter, broader wings ( Pteronotus macleayii, Pteronotus quadridens, Mormoops blainvillii) foraged most in edge habitats, but differed in timing of peak activity. P. macleayii and M. blainvillii differed in diet, but low sample size precluded diet analysis for P. quadridens. High-intensity echolocating species that used longer, more narrowband signals and had longer, narrower wings ( Molossus molossus, Tadarida brasiliensis) foraged most in open areas and differed in diet from the other species. Two disparate species were most active in clutter (dense vegetation). Pteronotus parnellii used high-duty-cycle echolocation apparently specialized for detecting fluttering targets in clutter. Macrotus waterhousii used low-intensity, broadband echolocation calls and presumably uses prey-generated sounds when foraging. These two species also differed in diet. Our data show that differences in morphology and echolocation behaviour coincide with differences in habitat use and diet, resulting in minimal overlap in resource use among species. [ABSTRACT FROM AUTHOR]

Migration is widespread among vertebrates, yet bat migration has received little attention and only in the recent decades has a better understanding of it been gained. Migration can cause significant changes in behaviour and physiology, due to increasing energy demands and aerodynamic constraints. Dietary shifts, for example, have been shown to occur in birds before onset of migration. For bats, it is not known if a change in diet occurs during migration, although breeding season-related dietary preference has been documented. It is known that a diet rich in fats and the accumulation of fat deposits do increase the flight range of migratory bats. Some bat species can be regarded as long-distance migrants, covering up to 2000 km between summer and winter roosting areas. Pipistrellus nathusii ( Vespertilionidae), a European long-distant migrant, travels each year along the Baltic Sea from north-eastern Europe to hibernate in central and southern Europe. This study presents data on the dietary habits of migrating Pipistrellus nathusii compared with those during the breeding season. We analysed faecal samples from bats on fall migration caught at the Ornithological Field Station in Pape, Latvia and from samples collected in North- Latvian summer roosts. We applied both morphological identification and molecular methods, as morphological methods also recognize life stages of prey and can contribute frequency data. The diets of bats on migration and breeding bats were similar, with Diptera and Lepidoptera comprising the major prey categories. However, certain prey groups could be explained by the different hunting habitats exploited during migration vs. summer residence. [ABSTRACT FROM AUTHOR]

Introduced species can exert disproportionately negative effects on island ecosystems, but their potential role as food for native consumers is poorly studied. Telfair's skinks are endemic omnivores living on Round Island, Mauritius, a globally significant site of biodiversity conservation. We aimed to determine the dietary diversity and key trophic interactions of Telfair's skinks, whether introduced species are frequently consumed, and if diet composition changes seasonally between male and female skinks. We used DNA metabarcoding of skink fecal samples to identify animals (COI) and plants (ITS2) consumed by skinks. There were 389 dietary presence counts belonging to 77 dietary taxa found across the 73 Telfair's skink fecal samples. Introduced taxa were cumulatively consumed more frequently than other categories, accounting for 49.4% of all detections, compared to cryptogenic (20.6%), native (20.6%), and endemic taxa (9.5%). The most frequently consumed introduced species was the ant, Pheidole megacephala, present in 40% of samples. Blue latan palm, Latania loddigesii, was the most frequently consumed endemic species, present in 33% of samples but was only detected in the dry season, when fruits are produced. We found a strong seasonal difference in diet composition explained by the presence of certain plant species solely or primarily in one season and a marked increase in the consumption of animal prey in the dry season. Male and female skinks consumed several taxa at different frequencies. These results present a valuable perspective on the role of introduced species in the trophic network of their invaded ecosystem. Both native and introduced species provide nutritional resources for skinks, and this may have management implications in the context of species conservation and island restoration. [ABSTRACT FROM AUTHOR]

Molecular sexing revealed an unexpectedly strong female bias in the sex ratio of pre-breeding European Storm Petrels (Hydrobates pelagicus), attracted to playback of conspecific calls during their northwards migration past SW Europe. This bias was consistent across seven years, ranging from 80.8% to 89.7% female (mean annual sex ratio ± SD = 85.5% female ± 4.1%). The sex ratio did not differ significantly from unity (i.e., 50% female) among (i) Storm Petrel chicks at a breeding colony in NW France, (ii) adults found dead on beaches in Southern Portugal, (iii) breeding birds attending nest burrows in the UK, captured by hand, and (iv) adults captured near a breeding colony in the UK using copies of the same sound recordings as used in Southern Europe, indicating that females are not inherently more strongly attracted to playback calls than males. A morphological discriminant function analysis failed to provide a good separation of the sexes, showing the importance of molecular sexing for this species. We found no sex difference in the seasonal or nocturnal timing of migration past Southern Europe, but there was a significant tendency for birds to be caught in sex-specific aggregations. The preponderance of females captured in Southern Europe suggests that the sexes may differ in migration route or in their colony-prospecting behaviour during migration, at sites far away from their natal colonies. Such differences in migration behaviour between males and females are poorly understood but have implications for the vulnerability of seabirds to pollution and environmental change at sea during the non-breeding season. [ABSTRACT FROM AUTHOR]

The analysis of food webs and their dynamics facilitates understanding of the mechanistic processes behind community ecology and ecosystem functions. Having accurate techniques for determining dietary ranges and components is critical for this endeavour. While visual analyses and early molecular approaches are highly labour intensive and often lack resolution, recent DNA-based approaches potentially provide more accurate methods for dietary studies. A suite of approaches have been used based on the identification of consumed species by characterization of DNA present in gut or faecal samples. In one approach, a standardized DNA region (DNA barcode) is PCR amplified, amplicons are sequenced and then compared to a reference database for identification. Initially, this involved sequencing clones from PCR products, and studies were limited in scale because of the costs and effort required. The recent development of next generation sequencing (NGS) has made this approach much more powerful, by allowing the direct characterization of dozens of samples with several thousand sequences per PCR product, and has the potential to reveal many consumed species simultaneously (DNA metabarcoding). Continual improvement of NGS technologies, on-going decreases in costs and current massive expansion of reference databases make this approach promising. Here we review the power and pitfalls of NGS diet methods. We present the critical factors to take into account when choosing or designing a suitable barcode. Then, we consider both technical and analytical aspects of NGS diet studies. Finally, we discuss the validation of data accuracy including the viability of producing quantitative data. [ABSTRACT FROM AUTHOR]

Little quantitative ecological information exists on the diets of most invertebrate feeding reptiles, particularly nocturnal or elusive species that are difficult to observe. In the UK and elsewhere, reptiles are legally required to be relocated before land development can proceed, but without knowledge of their dietary requirements, the suitability of receptor sites cannot be known. Here, we tested the ability of non-invasive DNA-based molecular diagnostics (454 pyrosequencing) to analyse reptile diets, with the specific aims of determining which earthworm species are exploited by slow worms (the legless lizard Anguis fragilis) and whether they feed on the deeper-living earthworm species that only come to the surface at night. Slow worm faecal samples from four different habitats were analysed using earthworm-specific PCR primers. We found that 86% of slow worms ( N = 80) had eaten earthworms. In lowland heath and marshy/acid grassland, Lumbricus rubellus, a surface-dwelling epigeic species, dominated slow worm diet. In two other habitats, riverside pasture and calciferous coarse grassland, diet was dominated by deeper-living anecic and endogeic species. We conclude that all species of earthworm are exploited by these reptiles and lack of specialization allows slow worms to thrive in a wide variety of habitats. Pyrosequencing of prey DNA in faeces showed promise as a practical, rapid and relatively inexpensive means of obtaining detailed and valuable ecological information on the diets of reptiles. [ABSTRACT FROM AUTHOR]

Rowson, B., Tattersfield, P. & Symondson, W. O. C. (2010). Phylogeny and biogeography of tropical carnivorous land-snails (Pulmonata: Streptaxoidea) with particular reference to East Africa and the Indian Ocean. - Zoologica Scripta, 40, 85-98. A phylogeny is presented for the speciose, near pan-tropical, carnivorous achatinoid land-snail superfamily Streptaxoidea inferred from DNA sequences (two nuclear and two mitochondrial regions) from 114 taxa from Africa, the Indian Ocean, Asia, South America and Europe. In all analyses, Streptaxidae are monophyletic, while the (two to six) previously recognised subfamilies are polyphyletic, as are several genus-level taxa including the most speciose genus Gulella, necessitating major taxonomic review. The Asian Diapheridae are sister to Streptaxidae, which forms several well-supported clades originating in a persistent basal polytomy. Divergence dating estimates, historical biogeography, and the fossil context suggest a Cretaceous origin of these families, but suggest Gondwanan vicariance predated most radiation. The basal polytomy dates to the Paleogene and may correspond to a rapid radiation in Africa. There is evidence for multiple Cenozoic dispersals followed by radiation, including at least two from Africa to South America, at least two from Africa to Asia and at least two from Africa to Madagascar, indicating Cenozoic turnover in tropical snail faunas. The endemic Seychelles and Mascarene streptaxid faunas each are composites of early Cenozoic lineages and more recent dispersals from Africa, with no direct evidence for an Asian origin as currently proposed. Peak streptaxid diversity in East Africa is explained by Neogene speciation among a large number of coexisting ancient lineages, a phenomenon most pronounced in the Eastern Arc-Coastal Forests centre of endemism. This includes Miocene diversification in Gulella, a primarily East and South-East African group which remains strikingly diverse even after unrelated ' Gulella' species are reclassified. [ABSTRACT FROM AUTHOR]

Earthworms play a major role in many aspects of soil fertility, food web ecology and ecosystem functioning, and hence are frequently the subjects of, for example, ecological and toxicological research. Our aim was to examine the genetic structure of common earthworm species, to identify cryptic lineages or species that may be distinct ecotypes or biotypes (and hence confound current research based upon morphotypes) and to try to explain the massive cryptic diversity that eventually emerged. We demonstrated that species such as Allolobophora chlorotica, Aporrectodea longa, Aporrectodea rosea and Lumbricus rubellus all comprise highly divergent lineages with species-level divergence at the mitochondrial cytochrome oxidase I (COI) gene. In Allo. chlorotica alone, we found 55 haplotypes for COI, with 35 of these being found in pink and 20 in green morph worms. There were no cases of the two colour morphs sharing COI haplotypes. Phylogenetic analyses of mitochondrial COI and 16S genes showed the presence of five highly divergent lineages, suggesting the presence of multiple cryptic species within Allo. chlorotica. There was no clear geographical pattern to lineage distribution and many populations were polymorphic for both mitochondrial DNA lineage and colour morph. Amplified fragment length polymorphism results, based on two primer combinations, were broadly congruent with mitochondrial DNA results with one significant exception. Despite showing over 14% divergence at COI, amplified fragment length polymorphism markers showed that the two green morph lineages may be interbreeding and therefore represent a single taxon. The cryptic diversity revealed by these results has profound consequences for all areas of earthworm research. [ABSTRACT FROM AUTHOR]

A molecular approach, using aphid-specific monoclonal antibodies, was used to test the hypothesis that alternative prey can affect predation on aphids by linyphiid spiders. These spiders locate their webs in cereal crops within microsites where prey density is high. Previous work demonstrated that of two subfamilies of Linyphiidae, one, the Linyphiinae, is web-dependent and makes its webs at sites where they were more likely to intercept flying insects plus those (principally aphids) falling from the crop above. The other, the Erigoninae, is less web-dependent, making its webs at ground level at sites with higher densities of ground-living detritivores, especially Collembola. The guts of the spiders were analysed to detect aphid proteins using enzyme-linked immunosorbent assay (ELISA). Female spiders were consuming more aphid than males of both subfamilies and female Linyphiinae were, as predicted, eating more aphid than female Erigoninae. Rates of predation on aphids by Linyphiinae were related to aphid density and were not affected by the availability of alternative prey. However, predation by the Erigoninae on aphids was significantly affected by Collembola density. Itinerant Linyphiinae, caught away from their webs, contained the same concentration of aphid in their guts as web-owners. However, nonweb-owning Erigoninae, living away from Collembola aggregations at web-sites, contained significantly higher concentrations of aphid. For both subfamilies there was evidence of a disproportionate increase in predation on aphids once Collembola populations had declined. It was concluded that nonaphid prey, by helping to maintain spiders in the field, can significantly affect predation on aphids. [ABSTRACT FROM AUTHOR]

Summary 1. Web-location cannot be designed simply to optimize foraging success, but must be a compromise between competing factors, including microhabitat parameters (physical structure, microclimate) and predator avoidance. 2. We tested the hypothesis that, despite these compromises, linyphiid spiders managed to locate their webs within microsites with enhanced prey resources. 3. We tested further the ‘equilibrium’ hypothesis that spiders, within a community of species, occupy different, relatively narrow niche axes and despite being generalists do not simply aggregate to the same resources. 4. In the first study of its kind, prey resources at web-sites were compared with those at matched non-web-sites with the same microhabitat properties. This was carried out for two subfamilies of Linyphiidae, one of which (Linyphiinae) is web-dependent and locates its web just above the ground, and the other (Erigoninae) is less web-dependent, locating its web on the ground and often hunting away from its web. We hypothesized that these subfamilies would locate their webs at microsites with enhanced densities of the prey most likely to be captured by their contrasting foraging strategies. We used spider weights as a surrogate for nutritional state, and compared spiders in webs with those without webs. 5. We found that even within a relatively uniform crop of wheat, spiders located their webs at microsites with greater prey resources. The most numerous prey, Collembola, were at significantly greater density at web-sites of both subfamilies of spider. However, there were significantly more Collembola at web-sites of the surface-hunting Erigoninae. By contrast, significantly more non-Collembola prey, especially aphids and Thysanoptera, were found at web-sites of the Linyphiinae, whose aerial webs might be expected to intercept such prey more efficiently. 6. These differences were also present at matched non-web-sites, suggesting that microsite selection by the spiders was determined by the spectra of prey present. Comparison of the weights of web-owning and non-web-owning Tenuiphantes tenuis (Linyphiinae) demonstrated the nutritional benefits of web-owning. 7. The results suggested that the spiders used a combination of web-location strategies at the microhabitat level, foraging behaviour and (known) high-intensity intraspecific competition, to exploit prey-rich microsites efficiently within fields in a dynamic manner. [ABSTRACT FROM AUTHOR]

Elucidating the diets of insect predators is important in basic and applied ecology, such as for improving the effectiveness of conservation biological control measures to promote natural enemies of crop pests. Here, we investigated the aphid diet of two common aphid predators in Central European agroecosystems, the native Coccinella septempunctata (Linnaeus) and the invasive Harmonia axyridis (Pallas; Coleoptera: Coccinellidae) by means of high throughput sequencing (HTS). For acquiring insights into diets of mobile flying insects at landscape scale minimizing trapping bias is important, which imposes methodological challenges for HTS. We therefore assessed the suitability of three field sampling methods (sticky traps, pan traps and hand-collection) as well as new aphid primers for identifying aphid prey consumption by coccinellids through HTS. The new aphid primers facilitate identification to species level in 75% of the European aphid genera investigated. Aphid primer specificity was high in silico and in vitro but low in environmental samples with the methods used, although this could be improved in future studies. For insect trapping we conclude that sticky traps are a suitable method in terms of minimizing sampling bias, contamination risk and trapping success, but compromise on DNA-recovery rate. The aphid diets of both field-captured ladybird species were dominated by Microlophium carnosum, the common nettle aphid. Another common prey was Sitobion avenae (cereal aphid), which got more often detected in C. septempunctata compared to H. axyridis. Around one third of the recovered aphid taxa were common crop pests. We conclude that sampling methodologies need constant revision but that our improved aphid primers offer currently one of the best solutions for broad screenings of coccinellid predation on aphids. [ABSTRACT FROM AUTHOR]

Among spiders, taxonomically the most diversified group of terrestrial predators, only a few species are stenophagous and feed on ants. The levels of stenophagy and ant-specialisation vary among such species. To investigate whether stenophagy is only a result of a local specialisation both fundamental and realised trophic niches need to be estimated. Here we investigated trophic niches in three closely-related spider species from the family Gnaphosidae (Callilepis nocturna, C. schuszteri, Nomisia exornata) with different levels of myrmecophagy. Acceptance experiments were used to estimate fundamental trophic niches and molecular methods to estimate realised trophic niches. For the latter two PCR primer sets were used as these can affect the niche breadth estimates. The general invertebrate ZBJ primers were not appropriate for detecting ant DNA as they revealed very few prey types, therefore ant-specific primers were used. The cut-off threshold for erroneous MOTUs was identified as 0.005% of the total number of valid sequences, at individual predator level it was 0.05%. The fundamental trophic niche of Callilepis species included mainly ants, while that of N. exornata included many different prey types. The realised trophic niche in Callilepis species was similar to its fundamental niche but in N. exornata the fundamental niche was wider than realised niche. The results show that Callilepis species are ant-eating (specialised) stenophagous predators, catching mainly Formicinae ants, while N. exornata is an ant-eating euryphagous predator catching mainly Myrmicinae ants. [ABSTRACT FROM AUTHOR]

Responses to the arguments on the spatial dynamics of predation by carabid beetles on the slugs Arion intermedius and Deroceras reticulatum. Reflection of demographic features on the predation; Spatial association between the slugs and P. melanarius; Impact of egg distribution on the distribution of slugs.

Predators that prey on dangerous species have evolved effective capture traits. In spiders, venom and silk represent distinct substances associated with prey capture. However, the use of such adaptations comes at a cost. Based on a possible trade-off, the use of only one effective capture mechanism should be optimized if a predator is to specialize on a single type of dangerous prey. We investigated hunting strategies in two Callilepis spp. and one Nomisia species, Nomisia exornata, closely related ant-eating spiders from the family Gnaphosidae. We hypothesized that specialized Callilepis spiders would adopt a more stereotyped capture strategy compared with less specialized Nomisia. We also expected that Callilepis would rely on only one subjugation mechanism. To test this, we compared their hunting efficacy and hunting strategies, with an emphasis on venom vs. silk utilization. Nomisia restrained ants with silk (then bit them), whereas Callilepis relied solely on its venom. This was also reflected in trophic traits connected with silk and venom utilization; Callilepis had larger venom glands than Nomisia, whereas Nomisia had more piriform silk glands than Callilepis. Callilepis was more effective because it subdued prey more quickly, presumably owing to ant-specific venom. Callilepis and Nomisia handled ants from two subfamilies with different degrees of success; Callilepis was more successful with Formicinae ants, whereas Nomisia handled Myrmicinae ants better. We show that sole reliance on venom allows Callilepis to be more efficient in overcoming ants than Nomisia, which uses both silk and venom. However, such specific adaptations might restrict specialized predators from exploiting alternative prey. [ABSTRACT FROM AUTHOR]

The article introduces a series of papers published within the issue, including one on the use of metabarcoding of gut contents to establish the diet of the brown shrimp, Crangon crangon, another on the prey of the wolf spider and a paper on associations between birds and feather mites determined by DNA metabarcoding.

Abstract: Dietary changes linked to the availability of anthropogenic food resources can have complex implications for species and ecosystems, especially when species are in decline. Here, we use recently developed primers targeting the ITS2 region of plants to characterize diet from faecal samples of four UK columbids, with particular focus on the European turtle dove (Streptopelia turtur), a rapidly declining obligate granivore. We examine dietary overlap between species (potential competition), associations with body condition in turtle doves and spatiotemporal variation in diet. We identified 143 taxonomic units, of which we classified 55% to species, another 34% to genus and the remaining 11% to family. We found significant dietary overlap between all columbid species, with the highest between turtle doves and stock doves (Columba oenas), then between turtle doves and woodpigeons (Columba palumbus). The lowest overlap was between woodpigeons and collared doves (Streptopelia decaocto). We show considerable change in columbid diets compared to previous studies, probably reflecting opportunistic foraging behaviour by columbids within a highly anthropogenically modified landscape, although our data for nonturtle doves should be considered preliminary. Nestling turtle doves in better condition had a higher dietary proportion of taxonomic units from natural arable plant species and a lower proportion of taxonomic units from anthropogenic food resources such as garden bird seed mixes and brassicas. This suggests that breeding ground conservation strategies for turtle doves should include provision of anthropogenic seeds for adults early in the breeding season, coupled with habitat rich in accessible seeds from arable plants once chicks have hatched. [ABSTRACT FROM AUTHOR]

Knowledge of the dietary choices and trophic niches of organisms is the key to understanding their roles in ecosystems. In seabird diet studies, prey identification is a difficult challenge, often yielding results with technique-specific biases. Additionally, sampling efforts are often not extensive enough to reveal intrapopulational variation. Immature animals, which may constitute up to 50% of a population, may occupy a significantly different trophic niche to more experienced birds, but this remains largely unexplored. We investigated the diet of Cory's shearwater ( Calonectris diomedea) from Selvagem Grande, an island located off the northwest African coast, collecting a total of 698 regurgitate samples over three consecutive breeding seasons. The diet was assessed using two complementary approaches for prey identification: conventional morphological analysis (using fish vertebrae, otoliths and cephalopod beaks) and DNA barcoding of the 16S r RNA mitochondrial gene, in cases where a positive identification could not be retrieved. Species assignments employed BLAST and distance-based methods, as well as direct optimization of the tree length based on unaligned sequences in POY. This method resulted in robust tree estimates and species assignments, showing its potential for DNA barcoding of stomach contents using hypervariable markers such as the 16S. The molecular approach increased taxonomic resolution and revealed an additional 17 taxa. Diet differed significantly according to breeding status, sex, breeding phase (prelaying and chick rearing) and year. Such direct evidence of trophic segregation within the same population has rarely been shown in seabirds and highlights the importance of including such variables in ecosystem-based management approaches. [ABSTRACT FROM AUTHOR]

Contamination pathways in complex food chains in soil ecosystems can be difficult to elucidate. Molecular analysis of predator gut content can, however, rapidly reveal previously unidentified trophic interactions between invertebrates and thereby uncover pathways of pollutant spread. Here, we measured concentrations of the toxic metals lead, cadmium and mercury in carabid beetle predators and their prey. Invertebrates were sampled at one control and four heavy metal-polluted sites to reveal the impact of diet composition and seasonal variation in prey availability on metal burden in carabids and metal transfer pathways through forest ecosystems. This is the first report, to our knowledge, of carabid diet composition based on PCR analysis of gut contents at the forest community level, rather than in cultivated fields. Extensive screening using group- and species-specific primers revealed that carabids ate primarily earthworms and slugs, as well as smaller numbers of woodlice and springtails. Metal concentrations in carabids correlated with seasonal changes in diet. Mercury accumulated in beetle predators more than in their slug prey. As earthworms, slugs and carabid beetles are the major prey of many birds and mammals, prey-predator transfer and associated toxicity are major risks at mercury-contaminated sites. Carabids may be useful bioindicators for assessing the impact of pollutants on soil ecosystems, as long as species and seasonal factors are taken into account. [ABSTRACT FROM AUTHOR]

The slugs of Britain and Ireland form a well-studied fauna of economic importance. They include many widespread European species that are introduced elsewhere (at least half of the 36 currently recorded British species are established in North America, for example). To test the contention that the British and Irish fauna consists of 36 species, and to verify the identity of each, a species delimitation study was conducted based on a geographically wide survey. Comparisons between mitochondrial DNA (COI, 16S), nuclear DNA (ITS-1) and morphology were investigated with reference to interspecific hybridisation. Species delimitation of the fauna produced a primary species hypothesis of 47 putative species. This was refined to a secondary species hypothesis of 44 species by integration with morphological and other data. Thirty six of these correspond to the known fauna (two species in Arion subgenus Carinarion were scarcely distinct and Arion (Mesarion) subfuscus consisted of two near-cryptic species). However, by the same criteria a further eight previously undetected species (22% of the fauna) are established in Britain and/or Ireland. Although overlooked, none are strictly morphologically cryptic, and some appear previously undescribed. Most of the additional species are probably accidentally introduced, and several are already widespread in Britain and Ireland (and thus perhaps elsewhere). At least three may be plant pests. Some evidence was found for interspecific hybridisation among the large Arion species (although not involving A. flagellus) and more unexpectedly in species pairs in Deroceras (Agriolimacidae) and Limacus (Limacidae). In the latter groups, introgression appears to have occurred in one direction only, with recently-invading lineages becoming common at the expense of long-established or native ones. The results show how even a well-studied, macroscopic fauna can be vulnerable to cryptic and undetected invasions and changes. [ABSTRACT FROM AUTHOR]

Reliable delimitation and identification of species is central not only to systematics, but also to studies of biodiversity, ecology and pest management. In the era of Internet-based biodiversity databases misidentifications are rapidly disseminated and may have far-reaching consequences. Leafhoppers from the genus Aphrodes ( Hemiptera, Cicadellidae) are common and abundant, but, nevertheless, they are still a taxonomically challenging group whose members are often assessed in ecological studies and are also potential vectors of plant diseases. Previous study has shown that the syntype series for A. aestuarina ( Edwards) includes also specimens of A. makarovi Zachvatkin and has suggested that misidentifications may be widespread in museum collections. We studied Aphrodes individuals collected from the U.K. and Slovenia in order to provide a more comprehensive analysis of this genus using multiple criteria. Combined work using male and female vibrational signals emitted during courtship, and a 600-bp fragment within the barcoding region of the COI mtDNA gene, provided validated specimens that we also used for morphometric study. Analyses confirmed A. aestuarina, A. bicincta, A. diminuta and A. makarovi as behaviourally, genetically and morphologically distinct species. Although any of these approaches could be used alone to distinguish between species, combining morphological and molecular approaches will help to improve reliability, especially when identifying females. Morphological investigation of validated individuals from the U.K. and Slovenia also revealed geographic differences within species. By combining several body and aedeagus morphological characters males can be reliably identified, however, morphological differences between species are, nevertheless, relatively small. By contrast, observed genetic distances between Aphrodes species are relatively large (4.2-7.0%). At about half of our collecting sites more than one Aphrodes species was found and A. makarovi was collected together with every other species, including A. aestuarina on tidal saltmarshes. Due to low morphological variation between syntopic congeners it is likely that many museum specimens of Aphrodes have been assigned to the wrong species and species identification in ecological and vector studies may also be questionable. [ABSTRACT FROM AUTHOR]

Predators can provide a valuable ecosystem service by suppressing crop pests. However, intraguild predation, where predators compete for the same prey resource whilst consuming each other, may destabilize population dynamics and increase the risk of pest outbreaks. Very little is known about intraguild predation in open fields or the strengths of trophic links between predators which may negatively affect pest control., We tested the null hypothesis that predation by the epigeal predator Pterostichus melanarius (Coleoptera: Carabidae) on different spiders is species-independent (proportional to density). A combination of population monitoring in winter wheat, molecular identification of juvenile spiders, molecular analysis of predator gut contents and a Monte Carlo simulation model were used to analyse prey choice., Pterostichus melanarius were pitfall-trapped over three months, and 622 individuals were screened for the remains of four spider species. Predation rates on spiders were 43·6% in June and 33·3% in August and showed clear evidence of prey choice., Predation on the web-dependent Tenuiphantes tenuis (Linyphiidae) was significantly greater than predicted from a random choice model, while predation on Bathyphantes gracilis (Linyphiidae) was significantly lower. The beetles may be selecting the most abundant species disproportionately (switching) or responding in some cases to spatial niche separation ( T. tenuis locate their webs marginally lower than B. gracilis). However, two itinerant hunters, Erigone spp. (Linyphiidae) and Pachygnatha degeeri (Tetragnathidae), were consumed in proportion to their density., Synthesis and applications. High levels of intraguild predation were revealed using molecular diagnostics. The gut analysis approach provided invaluable data that will inform the future design of appropriate pest management and integrated farming strategies that encourage these predators. The data showed strong evidence of prey choice. Managers can, however, probably encourage high densities of all these known aphid predators (spiders and carabids) because disproportionately high rates of predation on the most common spiders at our field sites ( T. tenuis) were not sufficient to prevent strong growth in the density of this species between June and August (adults increased × 1·6 and juveniles × 8·6). Such work is essential if we are to reveal the processes behind functional biodiversity in crops. [ABSTRACT FROM AUTHOR]

Biological control by conservation of native natural enemies can, at its best, reduce the need for pesticides and prevent detrimental effects upon the environment. The present study investigated the role of ground-active generalist predators as natural enemies of two tortricid pests in apple orchards., Predation rates were compared on the well established codling moth Cydia pomonella and the emerging oriental fruit moth Grapholita molesta, which has recently switched hosts to apples., The present study hypothesized that the ground-active predators consumed the two tortricid pests in significant numbers without preference, and attacked the pests at different developmental stages., Using diagnostic polymerase chain reaction on the gut contents of field-caught ground-active predators, no difference in predation rates was found on these two pests. Spiders were the most efficient predators of emergent adult moths in spring, whereas the carabid beetles, feeding on diapausing larvae, were important in the autumn., The temporal complementarity between spiders and carabid beetles, attacking different stages of the pests at different times of year, highlights the need for diverse predator assemblages to optimize biological control. [ABSTRACT FROM AUTHOR]

Primers were developed for leafhoppers of the genus Aphrodes amplifying 84-244 bp fragments of the mitochondrial cytochrome oxidase subunit I gene. DNA was extracted from legs of over 100-year-old archived museum specimens, amplified and sequenced. The fragments contain sufficient variation to unequivocally identify the different species. The majority of the analysed museum specimens, including three specimens of the syntype series for the UK endemic species A. aestuarina (Edwards 1908), were found to have been assigned to the wrong species. This work clearly underlines the need to validate museum specimens using molecular methods where identity is in doubt, based on reliable standards for species discrimination. [ABSTRACT FROM AUTHOR]

Vibrational signalling is a widespread form of animal communication and, in the form of sexual communication, has been generally regarded as inherently short-range and a private communication channel, free from eavesdropping by generalist predators. A combination of fieldwork and laboratory experiments was used to test the hypothesis that predators can intercept and exploit such signals. First, we developed and characterized PCR primers specific for leafhoppers of the genus Aphrodes and specifically for the species Aphrodes makarovi. Spiders were collected from sites where leafhoppers were present and screened with these primers to establish which spider species were significant predators of this species during the mating period of these leafhoppers. Analysis using PCR of the gut contents of tangle-web spiders, Enoplognatha ovata (Theridiidae), showed that they consume leafhoppers in the field at a greater rate when signalling adults were present than when nymphs were dominant, suggesting that the spiders were using these vibrations signals to find their prey. Playback and microcosm experiments then showed that E. ovata can use the vibrational signals of male leafhoppers as a cue during foraging and, as a result, killed significantly more male than female A. makarovi. Our results show, for the first time, that arthropod predators can exploit prey vibrational communication to obtain information about prey availability and use this information to locate and capture prey. This may be a widespread mechanism for prey location, one that is likely to be a major unrecognized driver of evolution in both predators and prey. [ABSTRACT FROM AUTHOR]

Species- and group-specific PCR primers were developed to study predation on pest and nonpest invertebrate species by generalist carabid predators in agroecosystems. To ensure the amplification of degraded DNA in predator gut samples, amplicons were designed to be less than 300 bp. Specificity of primers was assessed by cross-amplification against a panel of target and nontarget invertebrate species. The new primers were combined with previously published primers for slugs and collembolla in multiplex reactions to simultaneously screen each predator for the presence of multiple prey. All prey species were detected in a screen of the gut contents of field-caught predators. [ABSTRACT FROM AUTHOR]

The carabid beetle Pterostichus melanarius is a major natural enemy of pests, such as aphids and slugs in agricultural systems. Earthworms are a dominant non-pest component of the diet of P. melanarius which help sustain the beetles during periods when the pest population is low or absent. In this study we wanted to test whether this predator exercises prey choice among different earthworm species or ecological groups. High levels of genetic diversity within morphological species of earthworm necessitated the development of primers that were specific not just to species but lineages and sub-lineages within species as well. Gut samples from beetles were analysed using multiplex-PCR and fluorescent-labelled primers. Calibratory feeding trials were undertaken to calculate median detection times for prey DNA following ingestion. Extensive testing demonstrated that the primers were species-specific, that detection periods were negatively related to amplicon size and that meal size had a highly significant effect on detection periods. Monte Carlo simulations showed that, in general, worms were being predated in proportion to their densities in the field with little evidence of prey choice, other than probable avoidance of the larger, deep-living species. There was no evidence that epigeic species were being taken preferentially in comparison with endogeic species. There was also no evidence that defensive secretions by Allolobophora chlorotica reduced predation pressure on this species by P. melanarius. We concluded that any management system that increases earthworm densities generally, regardless of component species, is likely to be optimal for increasing numbers of this beneficial beetle predator. [ABSTRACT FROM AUTHOR]

In agricultural systems, polyphagous beetles and spiders are abundant components of the beneficial arthropod community. Although data on the dietary ranges of these groups is increasing, remarkably little is understood regarding how individuals interact with their prey at small spatial scales. We demonstrate the utility of a spatially-explicit network model that integrates predator behaviour using predator-prey co-occurrences. Three co-occurrence matrices, one each for June, July and August, were generated using Vortis suction sample data collected from an 80 point grid imposed on a field of winter wheat. Heuristic predator-prey linkages, based on positive spatial co-occurrence, were imposed on these three matrices to create networks. It was found that primary consumers were highly aggregated and showed a strong tendency to co-occur. This contrasted with patterns of predator–predator or predator–prey co-occurrences that either aggregated to their prey or were weak and more scattered. These patterns could not be explained by either competition for resources or body size differences. Procrustean methods indicated that the networks were temporally dynamic, consistently achieving rates of turnover >60%. A negative relationship was found between decreasing predator–prey co-occurrence in the network and the number of prey positives in the guts of those predators. For large polyphagous beetles, the closer they were to their prey at the field scale, the more likely they were to have eaten them. This simple underlying relationship suggests that spatial co-occurrence networks can be used to predict feeding behaviour and could make a valuable contribution to food web structuring. [ABSTRACT FROM AUTHOR]

1. Polyphagous predators, such as spiders and beetles, perform a fundamental ecosystem service as regulators of agricultural pests, particularly aphids. They are most effective when they colonize the crop before the pest has reached its exponential growth phase. However, this is also when predators find themselves in a state of near-starvation. 2. Predator numbers can be enhanced by applications of different types of organic matter, but the mechanism is not clearly understood. One hypothesis is that compost applied to the field may introduce a new detrital food chain to maintain predators until the pest arrives, but this may also be detrimental to effective pest control, fostering a surplus of alternative prey and causing a switch away from the pest. To elucidate these possible outcomes, we report on the use of within-field compost applications on aphids and their predators, presenting 4 years of field-scale manipulations. 3. We found both direct and indirect links between compost, aphids and predators. In years when compost-treated plots had significantly higher numbers of predators, aphids were in significantly lower numbers than in plots without compost. Conversely, when there was a lack of response by predators, aphid numbers showed similar trends in all treatments. 4. In all years, alternative prey responded strongly to compost application and did not fluctuate at the level shown by predators, suggesting that these two prey groups were decoupled. Instead, the predicted positive feedback of compost on predators numbers was either weak or absent. 5. Synthesis and applications. The effect of compost on aphids clearly requires further practical refinement if it is to provide constant pest suppression, making it difficult to provide specific management recommendations at this stage. In the short term, compost application may not always confer immediate benefits in terms of pest control alone but this must be set against other better known benefits (moisture retention, nutrients). In the long term, experiments measuring the full trophic pathway are needed to unravel the effects of organic matter type, application time and the siting of compost relative to the crop in order to optimise pest suppression potential. [ABSTRACT FROM AUTHOR]

Spatially explicit predator–prey interactions can alter the predatory potential of natural enemies augmented through conservation biological control. To test hypotheses regarding such interactions and predatory efficiency, we used a combination of molecular techniques and mark–release–recapture to study the foraging behaviour of a generalist carabid predator, Poecilus cupreus, in response to spatial patterns of its cereal aphid prey ( Metapolophium dirhodum and Sitobion avenae). Beetle and aphid numbers were measured across two grids of sampling locations, within which aphid spatial pattern had been manipulated to generate patchy and more homogenous distributions. Aphid consumption was measured by enzyme-linked immunosorbent assays (ELISA) of beetle gut contents, using an aphid-specific monoclonal antibody. Movement and distribution patterns suggest that P. cupreus does not aggregate at, nor instigate prey-taxis within, aphid patches. However, more than two-thirds of the 2169 P. cupreus tested by ELISA had consumed aphids and the proportion of beetles containing aphid proteins was positively related to aphid density. Against expectation, the proportion of predators feeding on aphids was greatest where prey were homogenously distributed, and this was attributed to the loss of partial refuges for prey in aphid patches. The functional value of this type of uniform foraging strategy is ideally suited to early colonization of the crop habitat, when aphid numbers are low, before populations build up and form strong spatial patterns. [ABSTRACT FROM AUTHOR]

Gut-content analyses using molecular techniques are an effective approach to quantifying predator–prey interactions. Predation is often assumed but scavenging is an equally likely route by which animal DNA enters the gut of a predator/scavenger. We used PCR (polymerase chain reaction) to detect scavenged material in predator gut homogenates. The rates at which DNA in decaying slugs (Mollusca: Pulmonata) and aphids (Homoptera: Aphididae) became undetectable were estimated. The detectability of DNA from both carrion types in the guts of the generalist predator Pterostichus melanarius (Coleoptera: Carabidae) was then determined. The effects of carrion age and weight, as well as beetle sex, on detection periods, were quantified. Laboratory trials measured prey preference of beetles between live and decaying prey. Further experiments measured, for the first time, feeding by P. melanarius on dead slugs and aphids directly in the field. In both field and laboratory, P. melanarius preferentially fed on dead prey if available, but preference changed as the prey became increasingly decayed. Disappearance rates for slug carrion in wheat fields and grasslands were estimated and P. melanarius was identified as the main scavenger. Comparison of the retention time for dead slugs in the field, with the detection period for decaying slug material in the guts of the predators, showed that PCR-based techniques are not able to distinguish between predated and scavenged food items. This could potentially lead to overestimation of the impact of predation on slugs (and other prey) by carabids. Possible implications of facultative scavenging by invertebrate predators for biocontrol and food-web research are discussed. [ABSTRACT FROM AUTHOR]

1. The spatial distribution of cereal aphids infesting a field of winter wheat during the population establishment, development and decline phases were studied using a field-scale grid of sampling locations.2. The distribution of two generalist predators,Pterostichus melanariusandP. madidus, were sampled contemporaneously.3. Using spatial analysis by distance indices (SADIE), spatial pattern in the aphid population, predator activity–density, predator hunger and aphid predation was detected and mapped.4. We tested the hypothesis that carabids and aphids were spatially associated with one another through predation. Aphid predation by individual beetles was detected using enzyme-linked immunosorbent assays (ELISA) and hunger was assessed by measurement of foregut weights.5. Initially, there was a strong spatial dissociation between aphids andP. melanariusactivity–density. While the aphid population was increasing there was a strong spatial association between aphids and bothP. melanariusandP. madidusactivity–density. During aphid population decline there was no measurable association between aphids and predatory activity–density.6. Predation of aphids was strongly locally associated with predator activity–density on all sample dates for both predator species, regardless of the association with aphid spatial pattern.7. Areas within the field most isolated fromP. melanariuspredation had the highest rates of aphid population increase.8. Although the proportion ofP. melanariusindividuals consuming aphids was much lower compared toP. madidus, it was concluded that it was a more effective biological control agent due to its comparative abundance.Journal of Animal Ecology(2005)doi: 10.1111/j.1365-2656.2005.00939.x [ABSTRACT FROM AUTHOR]

Summary: 1. An explicitly spatial sampling approach was employed to test the null hypothesis that the predation on slugs by the carabid beetle Pterostichus melanarius (Illiger) was opportunistic. 2. The beetles and slugs were sampled across a nested series of grids of sampling points, in a field of winter wheat during June and July 1997. 3. The spatial distribution of all slugs in June was found to change with the scale of the sampling grid, from random on the 0.25 m scale, through aggregation at 1 m, to random at 4 m. At the highest scale of 16 m, the slugs were significantly spatially aggregated. 4. The distribution of beetles in June was also spatially dynamic, with randomness observed at the 4 m and 8 m scales. At 16 m, significant aggregation was observed. 5. The dynamic distributions of slugs and beetles, at 16 m, were found not to be associated with, and thus were not determined by, soil or crop factors. 6. Comparison of slug and beetle populations showed, however, that the distributions at 16 m were dynamically associated with each other. In June where there were many slugs there were also many carabids, whilst in July where there were many carabids there were few slugs. 7. Approximately 11% of the beetles sampled across the 16 m grid in June and July were found to have ingested slug protein, following intensive enzyme-linked immunosorbent assay (ELISA) testing. 8. The spatial distribution of these slug-positive beetles was significantly associated with the distribution of the larger slug classes, over 25 mg. Where there were many large slugs in June there were many slug-positive beetles. Conversely, in July few large slugs were found where there were many slug-positive beetles. 9. Parametric analysis revealed that these changes in the large slug class, at each sampling point between June and July (growth), were negatively related to the local numbers of slug-positive beetles, and that growth declined as the local numbers of beetles increased. 10. ... [ABSTRACT FROM AUTHOR]

Simple Summary: Anoscopus leafhoppers are a group of plant-feeding bugs that can be found in a range of grassland habitats. There are seven recognized species in the UK, some of which are difficult to tell apart. One species, Anoscopus duffieldi, has only been found at a single site, an RSPB (Royal Society for the Protection of Birds) reserve at Dungeness in Kent. As Anoscopus leafhoppers can be quite variable in colour and pattern, and in the structure of their genitalia, our aim was to establish, using DNA, whether this 'species' is unique or whether it is simply a variant of one of the other species. If it is unique, found nowhere else, it should be afforded special protection. Samples of all UK species, as well as another from the Czech Republic, were collected from the field, and two genes were examined. The DNA sequences showed that three species, A. duffieldi, Anoscopus albifrons and Anoscopus limicola were so closely related that they should probably be considered a single species. However, A. duffieldi are distinctive in that they live only in one area of vegetated shingle. We suggest that, until other evidence is forthcoming, A. duffieldi could be considered a locally adapted subspecies of scientific interest. The subfamily Aphrodinae (Hemiptera: Cicadellidae) contains ~33 species in Europe within four genera. Species in two genera in particular, Aphrodes and Anoscopus, have proved to be difficult to distinguish morphologically. Our aim was to determine the status of the putative species Anoscopus duffieldi, found only on the RSPB Nature Reserve at Dungeness, Kent, a possible rare UK endemic. DNA from samples of all seven UK Anoscopus species (plus Anoscopusalpinus from the Czech Republic) were sequenced using parts of the mitochondrial cytochrome oxidase I and 16S rRNA genes. Bayesian inference phylogenies were created. Specimens of each species clustered into monophyletic groups, except for Anoscopusalbifrons, A. duffieldi and Anoscopuslimicola. Two A. albifrons specimens grouped with A. duffieldi repeatedly with strong support, and the remaining A. albifrons clustered within A. limicola. Genetic distances suggest that A. albifrons and A. limicola are a single interbreeding population (0% divergence), while A. albifrons and A. duffieldi diverged by only 0.28%. Shared haplotypes between A. albifrons, A. limicola and A. duffieldi strongly suggest interbreeding, although misidentification may also explain these topologies. However, all A. duffieldi clustered together in the trees. A conservative approach might be to treat A. duffieldi, until other evidence is forthcoming, as a possible endemic subspecies. [ABSTRACT FROM AUTHOR]

In Europe, the German Bight is one of the most important non-breeding areas for protected red-throated divers (Gavia stellata). It is unclear what attracts the birds to this area, especially as the food composition of seabirds outside the breeding season is notoriously difficult to study. To obtain information on prey species composition of red-throated divers in this area, faecal samples from 34 birds caught alive were analysed using DNA metabarcoding. Prey DNA was detected in 85% of the samples with a mean number of 4.2 ± 0.7 taxa per sample (n = 29). Altogether, we found a broad prey spectrum with 19 fish taxa from 13 families dominated by five groups: clupeids, mackerel, gadoids, flatfish and sand lances with clupeids being the most frequently detected prey. Our results indicate that red-throated divers are generalist opportunistic feeders in the German Bight, but pelagic schooling fish that aggregate at frontal zones and have a high energetic value might be favoured. Atlantic mackerel appears to be a more important prey for red-throated divers in this area than previously thought. The precision achievable using metabarcoding has revealed a number of prey species that are consumed by red-throated divers in the German Bight, which helps to explain the selection of this area by divers in winter and spring. [ABSTRACT FROM AUTHOR]

True predators are characterised by capturing a number of prey items during their lifetime and by being generalists. Some true predators are facultative specialists, but very few species are stenophagous specialists that catch only a few closely related prey types. A monophagous true predator that would exploit a single prey species has not been discovered yet. Representatives of the spider family Ammoxenidae have been reported to have evolved to only catch termites. Here we tested the hypothesis that Ammoxenus amphalodes is a monophagous termite-eater capturing only Hodotermes mossambicus. We studied the trophic niche of A. amphalodes by means of molecular analysis of the gut contents using Next Generation Sequencing. We investigated their willingness to accept alternative prey and observed their specific predatory behaviour and prey capture efficiency. We found all of the 1.4 million sequences were H. mossambicus. In the laboratory A. amphalodes did not accept any other prey, including other termite species. The spiders attacked the lateral side of the thorax of termites and immobilised them within 1 min. The paralysis efficiency was independent of predator:prey size ratio. The results strongly indicate that A. amphalodes is a monophagous prey specialist, specifically adapted to feed on H. mossambicus. [ABSTRACT FROM AUTHOR]