Although the practice of cultivating mass fl owering crops (MFCs) is seen as a means of counteracting the widespread decline of insect pollinators, no study to-date has evaluated whether the increased pollinator abundance associated with MFCs infl uences pollinator visits to assemblages of adjacent native, non-crop plant species (pollinator ' spill-over ' ). In this study we quantifi ed bumblebee abundance along hedgerow transects in MFC (fi eld bean) vs non-MFC (wheat) margins. Surveys were conducted on east and west-facing margins twice daily (morning and afternoon) for three or four days during the main MFC fl owering period (June) over four years involving paired bean and wheat fi elds in Devon and Cornwall, southwest England. Although no single bumblebee species showed any consistent change in relative frequency, when taken across all years and bumblebee species combined, we observed twice as many bumblebees visiting fl owers adjacent to the MFC. However, when we compared bumblebee activity along hedgerow transects two weeks after bean fl owering, there was no diff erence between crops. We thus conclude that although there is evidence for pollinator spillover from the bean MFC to nearby semi-natural habitat, the eff ect is comparatively short lived and not specifi c to any single bumblebee species. We also suggest that while pollinator services to native plant species might be briefl y enhanced by MFC cultivation, the possible repercussions of bumblebee spill-over from MFCs to semi-natural habitats and the other pollinator groups they support should be evaluated before MFC cultivation is considered as a means of conserving declining bumblebee populations.

Identifying and understanding predator diets is of high importance in biological conservation. This is particularly true for the introduction, establishment and maintenance of predator populations in newly created or modified ecological communities, such as translocation sites or restored habitats. Conservation status of predators may not permit captive feeding trials or intrusive gut-content methods, so non-intrusive diet assessment is required, such as faecal analysis. However, prey such as earthworms leave no morphological clues suitable for accurately discriminating between species consumed through visual faecal analysis. This study uses non-intrusive molecular methods on earthworm DNA extracted from the faeces of the carnivorous land snail Powelliphanta patrickensis to identify its earthworm diet and any seasonal trends. Data from 454-pyrosequencing revealed earthworm DNA in all samples (n = 60). Sequences were compared to a DNA library created from published and unpublished studies of New Zealand's endemic earthworms and online databases. Unidentified earthworm sequences were clustered into molecular operational taxonomic units (MOTUs). Twenty-six MOTUs were identified, 17 of which matched the library, whereas nine did not. Similarity indices indicate that there were seasonal differences (P < 0.05) in the earthworm communities represented in the summer and the winter diets. This study highlights the importance of utilising the vast body of data generated by pyrosequencing to investigate potential temporal diet shifts in protected species. The method described here is widely applicable to a wide range of predatory species of conservation interest and can further inform habitat restoration and relocation programmes to optimize the long-term survival of the target species.

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The use of DNA data is ubiquitous across animal sciences. DNA may be obtained from an organism for a myriad of reasons including identification and distinction between cryptic species, sex identification, comparisons of different morphocryptic genotypes or assessments of relatedness between organisms prior to a behavioural study. DNA should be obtained while minimizing the impact on the fitness, behaviour or welfare of the subject being tested, as this can bias experimental results and cause long-lasting effects on wild animals. Furthermore, minimizing impact on experimental animals is a key Refinement principle within the ‘3Rs’ framework which aims to ensure that animal welfare during experimentation is optimised. The term ‘non-invasive DNA sampling’ has been defined to indicate collection methods that do not require capture or cause disturbance to the animal, including any effects on behaviour or fitness. In practice this is not always the case, as the term ‘non-invasive’ is commonly used in the literature to describe studies where animals are restrained or subjected to aversive procedures. We reviewed the non-invasive DNA sampling literature for the past six years (380 papers published in 2013-2018) and uncovered the existence of a significant gap between the current use of this terminology (i.e. ‘non-invasive DNA sampling’) and its original definition. We show that 58% of the reviewed papers did not comply with the original definition. We discuss the main experimental and ethical issues surrounding the potential confusion or misuse of the phrase ‘non-invasive DNA sampling’ in the current literature and provide potential solutions. In addition, we introduce the terms ‘non-disruptive’ and ‘minimally disruptive’ DNA sampling, to indicate methods that eliminate or minimise impacts not on the physical integrity/structure of the animal, but on its behaviour, fitness and welfare, which in the literature reviewed corresponds to the situation for which an accurate term is clearly missing. Furthermore, we outline when these methods are appropriate to use.