Vinyl Ether/Tetrazine Pair for the Traceless Release of Alcohols in Cells

The cleavage of a protecting group from a protein or drug under bioorthogonal conditions enables accurate spatiotemporal control over protein or drug activity. Disclosed herein is that vinyl ethers serve as protecting groups for alcohol-containing molecules and as reagents for bioorthogonal bond-cleavage reactions. A vinyl ether moiety was installed in a range of molecules, including amino acids, a monosaccharide, a fluorophore, and an analogue of the cytotoxic drug duocarmycin. Tetrazine-mediated decaging proceeded under biocompatible conditions with good yields and reasonable kinetics. Importantly, the nontoxic, vinyl ether duocarmycin double prodrug was successfully decaged in live cells to reinstate cytotoxicity. This bioorthogonal reaction presents broad applicability and may be suitable for in vivo applications.European Commission; China Scholarship Council; FCT Portugal; MINECO (CTQ2015-70524-R and RYC-2013-14706); EPSRC; Royal Society; European Research Council (TagIt

EO9 (Apaziquone): from the clinic to the laboratory and back again

EO9 (Apaziquone) is a bioreductive drug that has a chequered history. It underwent clinical trial but failed to show activity in phase II clinical trials when administered i.v. Poor drug delivery to tumours caused by a combination of rapid pharmacokinetic elimination and poor penetration through avascular tissue were the major factors responsible for EO9’s poor efficacy. Based upon an understanding of why EO9 failed, a further clinical trial against patients with superficial transitional cell carcinoma of the bladder was conducted. The rationale for this was that intravesical administration directly into the bladder would circumvent the drug delivery problem, and any drug reaching the blood supply would be rapidly cleared thereby reducing the risk of systemic exposure. EO9 was well tolerated, and clinical activity against marker lesions was recorded in both phase I and II clinical trials. This article charts the pharmacological history of EO9 and discusses the potential implications that ‘the EO9 story’ has for the development of other loco-regional therapies

The trophic ecology of non-native ants on Round Island, Mauritius

Summary Non-native ants are implicated in the demise of native species around the world, though their trophic ecology remains poorly understood. Non-native ants have invaded Round Island, a globally significant site of biodiversity conservation located 21 km north-east of Mauritius in the Indian Ocean, but it is unclear how they are affecting the unique ecological community found there. To reveal their potential impact, I conducted a meta-analysis into the effects of non-native ants on animal community diversity in relatively undisturbed areas around the world, showing that non-native ants drive diversity declines in local animal communities by approximately 50 % on average (Chapter 2). I then examined the ecological role of non-native ants on Round Island specifically and first determined, using dietary DNA metabarcoding, whether an abundant native omnivore, Telfair’s skink, consumed non-native ants. Skinks do consume ants, though it was unclear to what degree these detections were deliberate or accidental (Chapter 3). I then identified the diet of the 12 most numerous non-native ant species on Round Island using dietary metabarcoding, revealing that all ant species showed unique generalist diet profiles and together consume over 150 species of animals and plants. The diet of the ant community was also driven by seasonal changes in food availability. This presents the first study to date detailing the diet of individual ants at the community level and that a community of generalist non-native species exhibit dietary niche separation (Chapter 4). I compared the diet of native skinks and centipedes with that of non-native ants, finding that skinks are not competing with ants for food, whilst centipedes are (Chapter 5). Overall, our results suggest, through five separate lines of evidence arising from the study, that non-native ants are having a significant impact on the Round Island ecosystem

Phylogenetic clustering of wingbeat frequency and flight-associated morphometrics across insect orders

Wingbeat frequency in insects is an important variable in aerodynamic and energetic analyses of insect flight and often is studied on a family‐ or species‐level basis. Meta‐analyses of these studies report order‐level patterns suggesting that flight strategy is moderately well conserved phylogenetically. Studies incorporated into these meta‐analyses, however, use variable methodologies across different temperatures, which may confound results and phylogenetic patterns. In the present study, a high‐speed camera is used to measure wingbeat frequency in a wide variety of species (n = 102) under controlled conditions aiming (i) to determine the validity of previous meta‐analyses showing phylogenetic clustering of flight strategy and (ii) to identify new evolutionary patterns between wingbeat frequency, body mass, wing area, wing length and wing loading at the order level. All flight‐associated morphometrics significantly affect wingbeat frequency. Linear models show that wing area explains the most amount of variation in wingbeat frequency (r2 = 0.59, P ≤ 0.001), whereas body mass explains the least (r2 = 0.09, P ≤ 0.01). A multiple regression model incorporating both body mass and wing area is the best overall predictor of wingbeat frequency (r2 = 0.84, P ≤ 0.001). Order‐level phylogenetic patterns across relationships are consistent with previous studies. Thus, the present study provides experimental validation of previous meta‐analyses and provides new insights into phylogenetically conserved flight strategies across insect orders

Density-independent prey choice, taxonomy, life history, and web characteristics determine the diet and biocontrol potential of spiders (Linyphiidae and Lycosidae) in cereal crops

Spiders are among the dominant invertebrate predators in agricultural systems and are significant regulators of insect pests. The precise dynamics of biocontrol of pests in the field are, however, poorly understood. This study investigates how density-independent prey choice, taxonomy, life stage, sex, and web characteristics affect spider diet and biocontrol. We collected spiders in four genera of Linyphiidae (i.e., Bathyphantes, Erigone, Tenuiphantes, and Microlinyphia), and individuals from the Lycosidae genus Pardosa, and their proximate prey communities from barley fields in Wales, UK between April and September 2018. We analyzed the gut contents of 300 individual spiders using DNA metabarcoding. From the 300 spiders screened, 89 prey taxa were identified from 45 families, including a wide range of pests and predators. Thrips were the dominant prey, present in over a third of the spiders sampled, but a type IV functional response appears to reduce their predation at peak abundances. Spider diets significantly differed based on web characteristics, but this depended on the genus and sex of the spider and it was not the principal separating factor in the trophic niches of linyphiids and lycosids. Diets significantly differed between spider genera and life stages, reflected in different propensities for intraguild predation and pest predation. Adult spiders predated a greater diversity of other predators, and juveniles predated a greater diversity of pests. Overall, Tenuiphantes spp. and Bathyphantes spp. exhibited the greatest individual potential for biocontrol of the greatest diversity of pest genera. The greater trophic niche complementarity of Pardosa spp. and Erigone spp., however, suggests that their complementary predation of different pests might be of greater overall benefit to biocontrol. Sustainable agriculture should aim to optimize conditions throughout the cropping cycle for effective biocontrol, prioritizing provision for a diversity of spiders which predate a complementary diversity of pest species

DNA metabarcoding reveals introduced species predominate in the diet of a threatened endemic omnivore, Telfair's skink (Leiolopisma telfairii)

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

Non‐native ants drive dramatic declines in animal community diversity: A meta‐analysis

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

The predator problem and PCR primers in molecular dietary analysis: Swamped or silenced; depth or breadth?

Dietary metabarcoding has vastly improved our ability to analyse the diets of animals, but it is hampered by a plethora of technical limitations including potentially reduced data output due to the disproportionate amplification of the DNA of the focal predator, here termed “the predator problem”. We review the various methods commonly used to overcome this problem, from deeper sequencing to exclusion of predator DNA during PCR, and how they may interfere with increasingly common multipredator-taxon studies. We suggest that multiprimer approaches with an emphasis on achieving both depth and breadth of prey detections may overcome the issue to some extent, although multitaxon studies require further consideration, as highlighted by an empirical example. We also review several alternative methods for reducing the prevalence of predator DNA that are conceptually promising but require additional empirical examination. The predator problem is a key constraint on molecular dietary analyses but, through this synthesis, we hope to guide researchers in overcoming this in an effective and pragmatic way

Temporal variation in spider trophic interactions is explained by the influence of weather on prey communities, web building and prey choice

1. 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. 2. 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. 3. 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. 4. 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

Money spider dietary choice in pre‐ and post‐harvest cereal crops using metabarcoding

Money spiders (Linyphiidae) are an important component of conservation biological control in cereal crops, but they rely on alternative prey when pests are not abundant, such as between cropping cycles. To optimally benefit from these generalist predators, prey choice dynamics must first be understood. Money spiders and their locally available prey were collected from cereal crops 2 weeks pre‐ and post‐harvest. Spider gut DNA was amplified with two novel metabarcoding primer pairs designed for spider dietary analysis, and sequenced. The combined general and spider‐exclusion primers successfully identified prey from 15 families in the guts of the 46 linyphiid spiders screened, whilst avoiding amplification of Erigone spp. The primers show promise for application to the diets of other spider families such as Agelenidae and Pholcidae. Distinct invertebrate communities were identified pre‐ and post‐harvest, and changes in spider diet and, to a lesser extent, prey choice reflected this. Spiders were found to consume one another more than expected, indicating their propensity towards intraguild predation, but also consumed common pest families. Changes in spider prey choice may redress prey community changes to maintain a consistent dietary intake. Consistent provision of alternative prey via permanent refugia should be considered to sustain effective conservation biocontrol