Raw data_Eitzinger et al_Effect of prey quality

Time series experiment with Lithobius spp. fed with either Collembola (Sinella curviseta), Lumbricidae (Lumbricus terrestris) or Diptera prey (Drosophila melanogaster). File includes data on succesful prey DNA detection, predator body mass before and after feeding and prey DNA quantity in predator guts. Missing data is indicated as N

Den materielle sannhets prinsipp - prinsippets vekt i dispositive saker i sivilprosessen

Avhandlingen vil tematisere spenningen som oppstår i balanseringen mellom prinsippet om materiell sannhet og andre prinsipper i de dispositive saker i sivilprosessen. Problemstillingen er om domstolens mulighet til å nå frem til et materielt riktig resultat i slike saker er underlagt for store begrensninger

Detection of flying insect prey DNA within dietary samples of different consumers (Linyphiidae, <i>Pardosa</i> spp.: whole body samples; <i>Nebria germari</i>, <i>N. jockischii</i>, <i>N. rufescens</i>, <i>Oreonebria castanea</i>: regurgitates; <i>Rhinolophus ferrumequinum</i>: faecal samples) when tested with the newly developed multiplex PCR systems FLY-1 and FLY-2.

<p>See main text for details on the multiplex PCR systems.</p><p>Detection of flying insect prey DNA within dietary samples of different consumers (Linyphiidae, <i>Pardosa</i> spp.: whole body samples; <i>Nebria germari</i>, <i>N. jockischii</i>, <i>N. rufescens</i>, <i>Oreonebria castanea</i>: regurgitates; <i>Rhinolophus ferrumequinum</i>: faecal samples) when tested with the newly developed multiplex PCR systems FLY-1 and FLY-2.</p

Group-Specific Multiplex PCR Detection Systems for the Identification of Flying Insect Prey

<div><p>The applicability of species-specific primers to study feeding interactions is restricted to those ecosystems where the targeted prey species occur. Therefore, group-specific primer pairs, targeting higher taxonomic levels, are often desired to investigate interactions in a range of habitats that do not share the same species but the same groups of prey. Such primers are also valuable to study the diet of generalist predators when next generation sequencing approaches cannot be applied beneficially. Moreover, due to the large range of prey consumed by generalists, it is impossible to investigate the breadth of their diet with species-specific primers, even if multiplexing them. However, only few group-specific primers are available to date and important groups of prey such as flying insects have rarely been targeted. Our aim was to fill this gap and develop group-specific primers suitable to detect and identify the DNA of common taxa of flying insects. The primers were combined in two multiplex PCR systems, which allow a time- and cost-effective screening of samples for DNA of the dipteran subsection Calyptratae (including Anthomyiidae, Calliphoridae, Muscidae), other common dipteran families (Phoridae, Syrphidae, Bibionidae, Chironomidae, Sciaridae, Tipulidae), three orders of flying insects (Hymenoptera, Lepidoptera, Plecoptera) and coniferous aphids within the genus <i>Cinara</i>. The two PCR assays were highly specific and sensitive and their suitability to detect prey was confirmed by testing field-collected dietary samples from arthropods and vertebrates. The PCR assays presented here allow targeting prey at higher taxonomic levels such as family or order and therefore improve our ability to assess (trophic) interactions with flying insects in terrestrial and aquatic habitats.</p></div

Gel image of PCR products amplified with the multiplex PCR systems FLY-1 and FLY-2 and separated with QIAxcel. FLY-1: Phoridae (Phor), Plecoptera (Plec), Tipulidae (Tipu), Sciaridae (Scia), Calyptratae (Calyp), artificial mix containing 300 double stranded (ds) templates per target.

<p>FLY-2: Syrphidae (Syrp), Hymenoptera (Hyme), Lepidoptera (Lepi), Bibionidae (Bibi), Chironomidae (Chiro), <i>Cinara</i> sp. (Cin.sp.), artificial mix with 250 ds templates per target. Note that an internal marker is run alongside each sample (15 and 3000 bp) and the scale on the left and right side, respectively, allows an estimation of fragment length. Sciaride may result in an additional weaker amplicon of ∼140 bp which is not interfering with another fragment in the system; the same applies for the long fragment that can be amplified from lepidopterans.</p

Evaluation of three molecular markers for identification of European primary parasitoids of cereal aphids and their hyperparasitoids

<div><p>Aphids are major pests of cereal crops and a suite of hymenopteran primary parasitoids play an important role in regulating their populations. However, hyperparasitoids may disrupt the biocontrol services provided by primary parasitoids. As such, understanding cereal aphid-primary parasitoid-hyperparasitoid interactions is vital for a reliable parasitoid-based control of cereal aphids. For this, the ability to identify the different primary and hyperparasitoid species is necessary. Unfortunately, this is often difficult due to a lack of morphologically diagnostic features. DNA sequence-based species identification of parasitoids can overcome these hurdles. However, comprehensive DNA sequence information is lacking for many of these groups, particularly for hyperparasitoids. Here we evaluate three genes [cytochrome <i>c</i> oxidase subunit I (COI), 16S ribosomal RNA (16S) and 18S ribosomal RNA (18S)] for their suitability to identify 24 species of primary parasitoids and 16 species of hyperparasitoids associated with European cereal aphids. To identify aphelinid primary parasitoid species and hyperparasitoids, we found 16S to be more suitable compared to COI sequences. In contrast, the Aphidiinae are best identified using COI due to better species-level resolution and a more comprehensive DNA sequence database compared to 16S. The 18S gene was better suited for group-specific identification of parasitoids, but did not provide resolution at the species level. Our results provide a DNA sequence database for cereal aphid primary parasitoids and their associated hyperparasitoids in Central Europe, which will allow further improvement of our understanding of cereal aphid-primary parasitoid-hyperparasitoid interactions in relation to aphid biological control.</p></div

Comparison of maximum within species distance (Max-WSD; X-axis) and minimum between species distance (Min-BSD, Y-axis) of non-Aphidiinae (aphelinid/hyperparasitoid) and Aphidiinae parasitoids for COI, 16S and 18S gene sequences.

<p>The percentages of species-pairs which have a Min-BSD smaller than the Max-WSD (“No barcoding gap”) and a Min-BSD smaller than the overall Max-WSD (“Min-BSD ≤ overall Max-WSD”) are shown. Points above the diagonal represent cases where the species pairs have barcoding gap.</p

Procrustes analysis of the results of the Correspondence analysis of the morphological and molecular data.

<p>Species' centroids are represented by triangles and the areas by circles. Closed shapes are molecular data; open shapes are morphological data. Early pioneer stage (A). late pioneer stage (B). Langtal (LT), Rotmoostal (RM), Gaisbergtal (GB). Species codes: Dip.hel (<i>Diplocephalus helleri</i>), Eri.tir (<i>Erigone tirolensis</i>), Agy.nig (<i>Agyneta nigripes</i>), Mec.pae (<i>Mecynargus paetulus</i>), Mug.var (<i>Mughiphantes variabilis</i>), Rob.aru (<i>Robertus arundineti</i>), Wal.wig (<i>Walckenaera vigilax</i>), Jan.mon (<i>Janetschekia monodon</i>), Eri.atr (<i>Erigone atra</i>), Ent.med (<i>Entelecara media</i>).</p

The two primer mixes for the multiplex PCR systems.

<p>The two primer mixes for the multiplex PCR systems.</p

Neighbour joining tree of the non-Aphidiinae parasitoids based on sequences of the 16S ribosomal RNA gene.

<p>Bootstrap values (≥ 70%) are indicated on branches. Species abbreviations see <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0177376#pone.0177376.g002" target="_blank">Fig 2</a>.</p