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Quantifying the growth of entomopathogenic fungi is crucial for understanding their virulence and pathogenic potential. Traditional methods for determining growth, such as biomass determination or colony growth area, are time-consuming and quantitatively and spatially limited in scope. In this study, we introduce a high-throughput method for rapidly measuring fungal growth using spectrophotometry in small-volume, liquid media cultures in 96-well microplates. Optical density (OD) changes were directly correlated with dry weight of samples for six isolates from three species of the genus Metarhizium to validate spectrophotometric growth measurements, and investigate species- and isolate-specific effects. We quantified fungal biomass from the microcultures by extracting, drying, and weighing mycelial mats. From the relationship established between OD and biomass, we generated standard curves for predicting biomass based on the OD values. The OD measurements clearly distinguished growth patterns among six isolates from three Metarhizium species. The logistic growth phase, as captured by the OD measurements, could be accurately assessed within a span of 80 h. Using isolates of M. acridum, M. brunneum, and M. guizhouense, this technique was demonstrated to be an effective, reproducible, and simple method for rapidly measuring filamentous fungal growth with high precision. This technique offers a valuable tool for studying the growth dynamics of entomopathogenic fungi and investigating the factors that influence their growth.

The One Health Priority Research Agenda on Antimicrobial Resistance (AMR) sets out for the first time the priorities for which the World Health Organization, the Food and Agriculture Organization of the United Nations, the United Nations Environment Programme, and the World Organisation for Animal Health – as leaders in the multilateral system on human, animal, plant, and environmental health – will advocate to promote research and investment in the response to AMR. The Research Agenda results from extensive stakeholder and expert engagement and was developed using a sound scientific methodology. The process identified major gaps in knowledge and evidence that require urgent scientific attention and resources. The document demonstrates how, by working together, we can effectively leverage our organizations’ respective resources and strengths in the multilateral system.

Here, we provide the genome sequence of a Leclercia adecarboxylata isolated from a screen of an environmental bacterial isolate library for resistance to the plant flavonoid berberine. We detected the colistin resistance gene mcr-9, located on an IncFII(pECLA) plasmid.

During 2021 and 2022 High Pathogenicity Avian Influenza (HPAI) killed thousands of wild birds across Europe and North America, suggesting a change in infection dynamics and a shift to new hosts, including seabirds. Northern Gannets (Morus bassanus) appeared especially severely impacted, but a detailed account of the data available is required to help understand how the virus spread across the metapopulation, and the ensuing demographic consequences. Accordingly, we analyse information on confirmed and suspected HPAIV outbreaks across most North Atlantic Gannet colonies and for the largest colony (Bass Rock, UK), provide impacts on population size, breeding success, and preliminary results on apparent adult survival and serology. Unusually high numbers of dead Gannets were first noted at colonies in Iceland during April 2022. Outbreaks in May occurred in many Scottish colonies, followed by colonies in Canada, Germany and Norway. By the end of June, outbreaks had occurred in colonies in Canada and the English Channel. Outbreaks in 12 UK and Ireland colonies appeared to follow a clockwise pattern with the last infected colonies recorded in late August/September. Unusually high mortality was recorded at 40 colonies (75% of global total colonies). Dead birds testing positive for HPAIV H5N1 were associated with 58% of these colonies. At Bass Rock, the number of occupied nest sites decreased by at least 71%, breeding success declined by ~66% compared to the long-term UK mean and the resighting of marked individuals suggested that apparent adult survival between 2021 and 2022 could have been substantially lower than the preceding 10-year average. Serological investigation detected antibodies specific to H5 in apparently healthy birds indicating that some Gannets recover from HPAIV infection. Further, most of these recovered birds had black irises, suggestive of a phenotypic indicator of previous infection. Untangling the impacts of HPAIV infection from other challenges face

Wild relatives of fruit trees (WRFT) are highly valued for food and tradable products by rural communities, especially in low-income countries and as such are a vital resource for tree improvement. During periods of conflict, a lack of support and protection by national authorities may make WRFT vulnerable. In Syria, WRFT are at risk of extinction due to the ongoing crisis, which has limited efforts to conserve and propagate these unique genetic resources. We collected information about the current status and key threats to WRFT in north west Syria from 50 agricultural experts using structured interviews. Our results show that many sites have experienced erosion, overgrazing and drought. To initiate in situ and ex situ conservation, the locations of WRFT exposed to deterioration were identified and mapped using GPS, and seeds from five genotypes per species were collected from each of ten species to be used for the establishment of an in situ and ex situ WRFT collection.

Using a citizen science approach, we identify a country-wide exposure to aerosolized spores of a human fungal pathogen, Aspergillus fumigatus, that has acquired resistance to the agricultural fungicide tebuconazole and first-line azole clinical antifungal drugs. Genomic analysis shows no distinction between resistant genotypes found in the environment and in patients, indicating that at least 40% of azole-resistant A. fumigatus infections are acquired from environmental exposures. Hotspots and coldspots of aerosolized azole-resistant spores were not stable between seasonal sampling periods. This suggests a high degree of atmospheric mixing resulting in an estimated per capita cumulative annual exposure of 21 days (±2.6). Because of the ubiquity of this measured exposure, it is imperative that we determine sources of azole-resistant A. fumigatus to reduce treatment failure in patients with aspergillosis.

Hawaiʻi is home to ʻoʻopu nākea (Awaous stamineus), a culturally significant, endemic, goby that exhibits an amphidromous life cycle characterized by a marine larval stage followed by post-larval recruitment to streams, where they live to become reproductive adults. However, it was recently suggested that their migration to the ocean might not be obligatory, as originally thought. Despite their importance in Hawaiian traditions and the ecology of Hawaiian freshwater ecosystems, we still lack a full understanding of their migratory patterns and life history due to the difficulties in determining the environmental migratory cues that set the timing and location of their migratory paths. This study examined environmental factors, such as mean annual rainfall, streamflow, and water chemistry, to determine if they play a role in whether A. stamineus spend their larval period in the ocean or their entire life cycle in freshwater streams. We sampled A. stamineus (n = 90) from three streams (Kahana, Kahaluʻu, and Waimānalo) on the island of Oʻahu, Hawaiʻi that represented the range of hydroclimatic gradient in wet-habitat conditions on the windward side of the island and characterized their migratory pattern using elemental analysis of sagittae, the largest pair of otoliths (calcareous ear structures). Based on otolith strontium:calcium and barium:calcium ratios, we determined if individuals spent their larval period in the ocean or the stream. We found that 100% of individuals displayed clear evidence of marine residence during their larval phase, regardless of the environmental factors the fish experienced. This study highlights the necessity of stream–ocean connectivity for the survival of A. stamineus and emphasizes the importance of stream-mouth conservation and management as it is a critical transition zone in stream–ocean–stream migratory pathways.

Wastewater-based epidemiology (WBE) for population-level surveillance of antimicrobial resistance (AMR) is gaining significant traction, but the impact of wastewater sampling methods on results is unclear. In this study, we characterized taxonomic and resistome differences between single-timepoint-grab and 24 h composites of wastewater influent from a large UK-based wastewater treatment work [WWTW (population equivalent: 223 435)]. We autosampled hourly influent grab samples (n=72) over three consecutive weekdays, and prepared additional 24 h composites (n=3) from respective grabs. For taxonomic profiling, metagenomic DNA was extracted from all samples and 16S rRNA gene sequencing was performed. One composite and six grabs from day 1 underwent metagenomic sequencing for metagenomic dissimilarity estimation and resistome profiling. Taxonomic abundances of phyla varied significantly across hourly grab samples but followed a repeating diurnal pattern for all 3 days. Hierarchical clustering grouped grab samples into four time periods dissimilar in both 16S rRNA gene-based profiles and metagenomic distances. 24H-composites resembled mean daily phyla abundances and showed low variability of taxonomic profiles. Of the 122 AMR gene families (AGFs) identified across all day 1 samples, single grab samples identified a median of six (IQR: 5–8) AGFs not seen in the composite. However, 36/36 of these hits were at lateral coverage <0.5 (median: 0.19; interquartile range: 0.16–0.22) and potential false positives. Conversely, the 24H-composite identified three AGFs not seen in any grab with higher lateral coverage (0.82; 0.55–0.84). Additionally, several clinically significant human AGFs (bla VIM, bla IMP, bla KPC) were intermittently or completely missed by grab sampling but captured by the 24 h composite. Wastewater influent undergoes significant taxonomic and resistome changes on short timescales potentially affecting interpretation of results based on sampling strategy. Grab sa

•1. Invasive non-native species and climate change are two of the greatest pressures facing freshwater communities; however, how they interact to impact ecosystem function remains poorly understood despite the potential for impacts on key functional behaviours, such as detritivory, which could have wide-reaching impacts. •2. We quantified the rates of detrital processing and survival of one U.K. native (Gammarus pulex) and two invasive non-native (Dikerogammarus villosus and Dikerogammarus haemobaphes) freshwater amphipod species, across three temperatures (8°C, 14°C, and 20°C), and three leaf diets of varying resource quality (oak, sycamore, and alder) in laboratory microcosms. We also compared amphipod survival across the different temperature and diet treatments. •3. Rates of detrital processing varied between the native and invasive non-native amphipod species, with native G. pulex having a faster processing rate than both invasive non-native species at the lower temperatures. However, as the temperature treatments increased, between-species differences decreased, while the effects of leaf diets became more apparent. Although the survival probability did not vary with amphipod species, amphipod survival was higher at lower temperatures and in treatments containing higher quality leaf diets. •4. We propose that the invasive non-native Dikerogammarus species will affect native communities through lower rates of detrital processing; however, this impact may change under predicted climatic warming and be increasingly similar to native amphipod species.

Plasmids enable the dissemination of antimicrobial resistance (AMR) in common Enterobacterales pathogens, representing a major public health challenge. However, the extent of plasmid sharing and evolution between Enterobacterales causing human infections and other niches remains unclear, including the emergence of resistance plasmids. Dense, unselected sampling is highly relevant to developing our understanding of plasmid epidemiology and designing appropriate interventions to limit the emergence and dissemination of plasmid-associated AMR. We established a geographically and temporally restricted collection of human bloodstream infection (BSI)-associated, livestock-associated (cattle, pig, poultry, and sheep faeces, farm soils) and wastewater treatment work (WwTW)-associated (influent, effluent, waterways upstream/downstream of effluent outlets) Enterobacterales. Isolates were collected between 2008-2020 from sites <60km apart in Oxfordshire, UK. Pangenome analysis of plasmid clusters revealed shared 'backbones', with phylogenies suggesting an intertwined ecology where well-conserved plasmid backbones carry diverse accessory functions, including AMR genes. Many plasmid 'backbones' were seen across species and niches, raising the possibility that plasmid movement between these followed by rapid accessory gene change could be relatively common. Overall, the signature of identical plasmid sharing is likely to be a highly transient one, implying that plasmid movement might be occurring at greater rates than previously estimated, raising a challenge for future genomic One Health studies.

The approach based on analysing wastewater to monitor drugs or pathogens in the population had been used for decades before it became widespread during the COVID 19 pandemic. We have asked experts located in a number of different nations to share their views on the potential and limitations of wastewater-based epidemiology in the future.

•Background: The COVID-19 pandemic showcased the power of genomic sequencing to tackle the emergence and spread of infectious diseases. However, metagenomic sequencing of total microbial RNAs in wastewater has the potential to assess multiple infectious diseases simultaneously and has yet to be explored. •Methods: A retrospective RNA-Seq epidemiological survey of 140 untreated composite wastewater samples was performed across urban (n = 112) and rural (n = 28) areas of Nagpur, Central India. Composite wastewater samples were prepared by pooling 422 individual grab samples collected prospectively from sewer lines of urban municipality zones and open drains of rural areas from 3rd February to 3rd April 2021, during the second COVID-19 wave in India. Samples were pre-processed and total RNA was extracted prior to genomic sequencing. •Findings: This is the first study that has utilised culture and/or probe-independent unbiased RNA-Seq to examine Indian wastewater samples. Our findings reveal the detection of zoonotic viruses including chikungunya, Jingmen tick and rabies viruses, which have not previously been reported in wastewater. SARS-CoV-2 was detectable in 83 locations (59%), with stark abundance variations observed between sampling sites. Hepatitis C virus was the most frequently detected infectious virus, identified in 113 locations and co-occurring 77 times with SARS-CoV-2; and both were more abundantly detected in rural areas than urban zones. Concurrent identification of segmented virus genomic fragments of influenza A virus, norovirus, and rotavirus was observed. Geographical differences were also observed for astrovirus, saffold virus, husavirus, and aichi virus that were more prevalent in urban samples, while the zoonotic viruses chikungunya and rabies, were more abundant in rural environments. •Interpretation: RNA-Seq can effectively detect multiple infectious diseases simultaneously, facilitating geographical and epidemiological surveys of endemic viruse

Antidepressants are one of the most commonly prescribed pharmaceuticals. Although they have been frequently detected in aquatic environments around the globe, little is known regarding their adverse effects on humans and aquatic organisms. Recently, an in vitro monoamine transporter inhibition assay was developed to detect transporter-inhibitory activities of antidepressants in wastewater in Japan. However, it was unclear which antidepressants were responsible for transporter-inhibitory activities in wastewater. Herein, the per capita consumption of 32 antidepressants, their excretion of unchanged parent compounds, per capita water consumption, removal rate during wastewater treatment processes, and potency values from the monoamine transporter inhibition assay were used to prioritize antidepressants of concern in effluent wastewater in England and Japan. In both countries, sertraline and O-desmethylvenlafaxine had the highest contribution to inhibitory activities against the human serotonin transporter (hSERT) and zebrafish SERT (zSERT), respectively. It was found that the antidepressants inhibited the zSERT more strongly than the hSERT. The inhibitory activities found against the zSERT in wastewater in England and Japan were higher than thresholds for abnormal behavior in fish. The antidepressants prioritized in this study provide insight into launching environmental monitoring and ecotoxicological studies of antidepressants.

Forest-based communities manage many risks to health and socio-economic welfare including the increasing threat of emerging zoonoses that are expected to disproportionately affect poor and marginalised groups, and further impair their precarious livelihoods, particularly in Low-and-Middle Income (LMIC) settings. Yet, there is a relative dearth of empirical research on the vulnerability and adaptation pathways of poor and marginalised groups facing emerging zoonoses. Drawing on a survey of 229 households and a series of key-informant interviews in the Western Ghats, we examine the factors affecting vulnerability of smallholder and tribal households to Kyasanur Forest Disease (KFD), an often-fatal tick-borne viral haemorrhagic fever endemic in south India. Specifically, we investigate how different socio-demographic and institutional factors interact to shape KFD vulnerability and the strategies employed by households to adapt to disease consequences. Although surveyed households generally perceived KFD as an important health issue in the study region, there was variability in concern about contracting the disease. Overall results showed that poor access to land (AOR = 0.373, 95% CI: 0.152–0.916), being at or below the poverty line (AOR = 0.253, 95% CI: 0.094–0.685) and being headed by an older person (AOR = 1.038, 95% CI: 1.006–1.071) were all significant determinants of perceived KFD vulnerability. Furthermore, KFD vulnerability is also modulated by important extra-household factors including proximity to private hospitals (AOR = 3.281, 95% CI: 1.220–8.820), main roads (AOR = 2.144, 95% CI: 1.215–3.783) and study location (AOR = 0.226, 95% CI: 0.690–0.743). Our findings highlight how homogenous characterisation of smallholder and tribal communities and the ‘techno-oriented’ approach of existing interventions may further marginalise the most vulnerable and exacerbate existing inequalities. These findings are important for designing context-specific and appropriate he

Globally, major efforts are being made to restore peatlands to maximise their resilience to anthropogenic climate change, which puts continuous pressure on peatland ecosystems and modifies the geography of the environmental envelope that underpins peatland functioning. A probable effect of climate change is reduction in the waterlogged conditions that are key to peatland formation and continued accumulation of carbon (C) in peat. C sequestration in peatlands arises from a delicate imbalance between primary production and decomposition, and microbial processes are potentially pivotal in regulating feedbacks between environmental change and the peatland C cycle. Increased soil temperature, caused by climate warming or disturbance of the natural vegetation cover and drainage, may result in reductions of long-term C storage via changes in microbial community composition and metabolic rates. Moreover, changes in water table depth alter the redox state and hence have broad consequences for microbial functions, including effects on fungal and bacterial communities especially methanogens and methanotrophs. This article is a perspective review of the effects of climate change and ecosystem restoration on peatland microbial communities and the implications for C sequestration and climate regulation. It is authored by peatland scientists, microbial ecologists, land managers and non-governmental organisations who were attendees at a series of three workshops held at The University of Manchester (UK) in 2019–2020. Our review suggests that the increase in methane flux sometimes observed when water tables are restored is predicated on the availability of labile carbon from vegetation and the absence of alternative terminal electron acceptors. Peatland microbial communities respond relatively rapidly to shifts in vegetation induced by climate change and subsequent changes in the quantity and quality of below-ground C substrate inputs. Other consequences of climate change that affec

The yellow mealworm, Tenebrio molitor, is currently one of the most important insect species produced for livestock feed and human consumption. High-density rearing conditions make the risk of disease and infections by parasitic symbionts a challenge in the mass production of these insects. However, certain symbionts are beneficial and should be favoured in order to promote healthy insect populations. Knowledge of parasitic symbionts and their management is essential for the insect rearing industry and its associated research. Here we review the documented microbial infectious agents, invertebrate parasites, and beneficial symbionts occurring in T. molitor. Furthermore, we discuss detection, prevention, and treatment methods for disease management in T. molitor production systems to inform future management and decision making in T. molitor rearing.

Trace elements are chemical contaminants spread in the environment by anthropogenic activities and threaten wildlife and human health. Many studies have investigated this contamination in apex raptors as sentinel birds. However, there is limited data for long-term biomonitoring of multiple trace elements in raptors. In the present study, we measured the concentrations of 14 essential and non-essential trace elements in the livers of the common buzzard (Buteo buteo) collected in the United Kingdom from 2001 to 2019 and investigated whether concentrations have changed during this period. In addition, we estimated the importance of selected variables for modelling element accumulations in tissues. •Except for cadmium, hepatic concentrations of harmful elements in most buzzards were lower than the biological significance level of each element. Hepatic concentrations of certain elements, including lead, cadmium, and arsenic, varied markedly seasonally within years. Their peak was in late winter and trough in late summer, except copper which showed an opposite seasonal pattern. In addition, lead in the liver consistently increased over time, whereas strontium showed a decreasing trend. Hepatic concentrations of cadmium, mercury, and chromium increased with age, whereas selenium and chromium were influenced by sex. Hepatic concentrations of arsenic and chromium also differed between different regions. •Overall, our samples showed a low risk of harmful effects of most elements compared to the thresholds reported in the literature. Seasonal fluctuation was an important descriptor of exposure, which might be related to the diet of the buzzard, the ecology of their prey, and human activities such as the use of lead shot for hunting. However, elucidating reasons for these observed trends needs further examination, and biomonitoring studies exploring the effects of variables such as age, sex, and seasonality are required.

Palaeolimnological records provide valuable information about how phytoplankton respond to long-term drivers of environmental change. Traditional palaeolimnological tools such as microfossils and pigments are restricted to taxa that leave sub-fossil remains, and a method that can be applied to the wider community is required. Sedimentary DNA (sedDNA), extracted from lake sediment cores, shows promise in palaeolimnology, but validation against data from long-term monitoring of lake water is necessary to enable its development as a reliable record of past phytoplankton communities. To address this need, 18S rRNA gene amplicon sequencing was carried out on lake sediments from a core collected from Esthwaite Water (English Lake District) spanning ~105 years. This sedDNA record was compared with concurrent long-term microscopy-based monitoring of phytoplankton in the surface water. Broadly comparable trends were observed between the datasets, with respect to the diversity and relative abundance and occurrence of chlorophytes, dinoflagellates, ochrophytes and bacillariophytes. Up to 20% of genera were successfully captured using both methods, and sedDNA revealed a previously undetected community of phytoplankton. These results suggest that sedDNA can be used as an effective record of past phytoplankton communities, at least over timescales of <100 years. However, a substantial proportion of genera identified by microscopy were not detected using sedDNA, highlighting the current limitations of the technique that require further development such as reference database coverage. The taphonomic processes which may affect its reliability, such as the extent and rate of deposition and DNA degradation, also require further research.

Invasive species are the second largest contributor to biodiversity loss and drivers of ecosystem change. Buffelgrass is a C4, perennial grass native to Africa and Asia that was widely introduced across tropical and subtropical rangelands as livestock forage. Buffelgrass reduces native biodiversity and ecosystem stability in its introduced range when it escapes livestock pastures. Understanding the ecology of insects associated with buffelgrass in its native range may provide an understanding of invasion processes and biological control opportunities where buffelgrass has become an invasive challenge. Here, we present the results of a six-year survey of herbivorous arthropods of a native buffelgrass population from Kenya. Buffelgrass was examined for externally and internally feeding insects of vegetative and reproductive tissues. We also categorised buffelgrass detritivores and parasitoids that may use buffelgrass herbivores as hosts. The samples were photographed and Sanger sequenced to identify them to the lowest possible taxonomic ranking. We collected information on sample abundances, phenologies, tissues consumed, and putative diet breadths. We identified 25 morphospecies representing seven orders and 16 families. The putative host plant specialisation was as high as 67% for Diptera. Phenological variation in herbivore presence correlated with seasonal rainfall and provided a guide for when to conduct follow-up biological control agent searches. The most abundant herbivore was a gall midge (Diptera: Cecidomyiidae) that inhabits buffelgrass culms. Additional research should focus on bringing this species into containment where host choice trials can be conducted to determine if it is truly monophagous and assess its impact on buffelgrass growth.

•1. During primary succession, the abundance of different species and their associated plant traits change over time. Understanding how plant traits linked to colonising and competitive abilities change through succession is important for determining whether community assembly can be predicted. Examining this across more than one taxon group can reveal if these patterns are generalisable. •2. Here, we investigated primary succession on bare chalk for a chronosequence spanning 33 years for two different taxa, vascular plants and bryophytes. We examined how abundance changed through succession, and how this related to species' colonising and competitive abilities, using relevant plant traits for each taxa. A zero-inflated beta regression model was used to investigate the effects of traits on both presence/absence and abundance-when-present of vascular plants and bryophytes. •3. Vascular plants with a larger specific leaf area were more likely to occur later in succession. Vascular plants, which were hemicryptophytes, wind dispersing and had a lower canopy height, were more likely to increase in abundance-when-present during succession. •4. Bryophytes with a larger spore diameter were more likely to occur later in succession. Shorter bryophytes with a greater frequency of sporophyte production had a higher abundance early in succession, representing their high colonising abilities. Whereas later in succession larger bryophytes, with a mat or weft life form and low sporophyte frequency were more abundant, indicating a shift towards greater competitive abilities. •5. Synthesis. This study has revealed different patterns for vascular plants and bryophytes regarding colonisation and changes in abundance through succession, and the associated traits linked to colonising and competitive abilities. Although some traits were found to influence abundance through succession for vascular plants, these were often contrary to the expected pattern representing the change from coloni

Color vision in insects is determined by signaling cascades, central to which are opsin proteins, resulting in sensitivity to light at different wavelengths. In certain insect groups, lineage-specific evolution of opsin genes, in terms of copy number, shifts in expression patterns, and functional amino acid substitutions, has resulted in changes in color vision with subsequent behavioral and niche adaptations. Lepidoptera are a fascinating model to address whether evolutionary change in opsin content and sequence evolution are associated with changes in vision phenotype. Until recently, the lack of high-quality genome data representing broad sampling across the lepidopteran phylogeny has greatly limited our ability to accurately address this question. Here, we annotate opsin genes in 219 lepidopteran genomes representing 33 families, reconstruct their evolutionary history, and analyze shifts in selective pressures and expression between genes and species. We discover 44 duplication events in opsin genes across ∼300 million years of lepidopteran evolution. While many duplication events are species or family specific, we find retention of an ancient long-wavelength-sensitive (LW) opsin duplication derived by retrotransposition within the speciose superfamily Noctuoidea (in the families Nolidae, Erebidae, and Noctuidae). This conserved LW retrogene shows life stage–specific expression suggesting visual sensitivities or other sensory functions specific to the early larval stage. This study provides a comprehensive order-wide view of opsin evolution across Lepidoptera, showcasing high rates of opsin duplications and changes in expression patterns.

The yellow mealworm (Tenebrio molitor) is a promising insect species for mass-rearing for the production of feed and food. In mass-production systems, insects may be exposed to abiotic stressors such as heat stress as well as potentially lethal pathogens. To ensure mass-reared T. molitor populations are healthy and productive there is a need to understand both the risks, and potential benefits of heat stress, on the fitness of insects and their susceptibility to pathogens. In this study, we investigated the effects of a short (2 h) or a long (14 h) heat stress (38 °C) exposure on the susceptibility and the immune responses of T. molitor larvae exposed to a fungal pathogen (Metarhizium brunneum). Larvae were exposed to the pathogen either immediately or five days after the heat stress treatments. The development of heat stressed larvae and their offspring was also assessed. A short heat stress immediately before exposure to M. brunneum increased the survival probability of T. molitor larvae, which correlated with increased antibacterial activity in the hemolymph. The exposure of larvae to short, or long heat stresses five days before pathogen exposure did not affect their survival, despite a temporary lowered body mass gain of heat stressed larvae. However, heat stressed larvae showed decreased hemocyte concentrations when exposed to M. brunneum. We also found an increased body weight in larval offspring of females that had been exposed to a short heat stress as larvae themselves. These findings demonstrate the importance of understanding the effects of heat stress in the long-term. The beneficial effects of heat stress on pathogen susceptibility in T. molitor and the negative effects on body mass gain are only transient, whereas negative effects on immune response (hemocyte concentrations) persist over an extended period.

African swine fever virus (ASFV) is known to be very stable and can remain infectious over long periods of time especially at low temperatures and within different matrices, particularly those containing animal-derived organic material. However, there are some gaps in our knowledge pertaining to the survivability and infectivity of ASFV in groundwater. This study aims to determine the stability and infectivity of the cell culture-adapted ASFV strain BA71V by plaque assay after incubation of the virus within river water samples at three different environmentally relevant temperatures (4 °C, 15 °C, and 21 °C) over the course of 42 days. The results from this study indicate that ASFV can remain stable and infectious when maintained at 4 °C in river water for more than 42 days, but as incubation temperatures are increased, the stability is reduced, and the virus is no longer able to form plaques after 28 days and 14 days, respectively, when stored at 15 °C and 21 °C. Characterizing the survivability of ASFV in groundwater can allow us to develop more appropriate inactivation and disinfection methods to support disease control and mitigate ASFV outbreaks.

Professor Alan Raybould was born and raised in Wolverhampton, in the West Midlands, United Kingdom. He attained a First Class Degree in Botany from the University of Manchester, followed by his PhD in Population Genetics at the University of Birmingham in 1989, researching population genetics of Spartina anglica. Alan began his scientific career at the Institute of Terrestrial Ecology at Furzebrook, Dorset, which later became part of the United Kingdom Centre for Ecology and Hydrology. During this period (1990–2001), he progressed from a post-doctoral research position to becoming the lead scientist in molecular ecology, studying gene-flows from genetically-modified crops to related wild plant populations.

•Introduction: There are concerns that antimicrobial usage (AMU) is driving an increase in multi-drug resistant (MDR) bacteria so treatment of microbial infections is becoming harder in humans and animals. The aim of this study was to evaluate factors, including usage, that affect antimicrobial resistance (AMR) on farm over time. •Methods: A population of 14 cattle, sheep and pig farms within a defined area of England were sampled three times over a year to collect data on AMR in faecal Enterobacterales flora; AMU; and husbandry or management practices. Ten pooled samples were collected at each visit, with each comprising of 10 pinches of fresh faeces. Up to 14 isolates per visit were whole genome sequenced to determine presence of AMR genes. •Results: Sheep farms had very low AMU in comparison to the other species and very few sheep isolates were genotypically resistant at any time point. AMR genes were detected persistently across pig farms at all visits, even on farms with low AMU, whereas AMR bacteria was consistently lower on cattle farms than pigs, even for those with comparably high AMU. MDR bacteria was also more commonly detected on pig farms than any other livestock species. •Discussion: The results may be explained by a complex combination of factors on pig farms including historic AMU; co-selection of AMR bacteria; variation in amounts of antimicrobials used between visits; potential persistence in environmental reservoirs of AMR bacteria; or importation of pigs with AMR microbiota from supplying farms. Pig farms may also be at increased risk of AMR due to the greater use of oral routes of group antimicrobial treatment, which were less targeted than cattle treatments; the latter mostly administered to individual animals. Also, farms which exhibited either increasing or decreasing trends of AMR across the study did not have corresponding trends in their AMU. Therefore, our results suggest that factors other than AMU on individual farms are important for p

In anuran amphibians (frogs and toads), evidence linking pollution to population declines is limited, in particular through impaired reproduction. Here we review the evidence for pollutant-induced alterations on reproductive endpoints in wild anurans with a particular focus on the application of non-destructive endpoints including on sex ratios, male reproductive phenotypes (data are too scarce for females) and reproductive outputs (reflective of mating success). Data evidencing alterations in sex ratio in wild anurans are scarce, however, both feminisation and masculinisation in response to pollution have been reported (seven studies). Male nuptial pad morphology and calling behaviour display high sensitivity to pollutant-exposure and are important features determining male breeding success, however there is considerable variation in these endpoints and inconsistencies in the responses of them to pollution are reported in wild anurans. Data for clutch size are insufficient to assess sensitivity to pollutants (five studies only). However, hatch success and offspring fitness (tadpole survival/development) are sensitive to pollution, with clear linkages to population stability. In conclusion, there are a wide range of non destructive measures with good potential for application to assess/monitor reproductive health in wild anurans, however, a greater understanding of pollutant effects on these endpoints is needed. There measures deserve wider application as they are relatively simple and inexpensive to implement, and as they can be applied non-destructively are widely applicable to our declining anuran populations.

We present a genome assembly from an individual male Eupithecia dodoneata (the Oak-tree Pug; Arthropoda; Insecta; Lepidoptera; Geometridae). The genome sequence is 353.7 megabases in span. Most of the assembly is scaffolded into 31 chromosomal pseudomolecules, including the assembled Z sex chromosome. The mitochondrial genome has also been assembled and is 15.3 kilobases in length.

Xylella fastidiosa is a xylem-limited plant pathogen infecting many crops globally and is the cause of the recent olive disease epidemic in Italy. One strategy proposed to mitigate losses is to replant susceptible crops with resistant varieties. Several genetic, biochemical and biophysical traits are associated to X. fastidiosa disease resistance. However, mechanisms underpinning resistance are poorly understood. We hypothesize that the susceptibility of olive cultivars to infection will correlate to xylem vessel diameters, with narrower vessels being resistant to air embolisms and having slower flow rates limiting pathogen spread. To test this, we scanned stems from four olive cultivars of varying susceptibility to X. fastidiosa using X-ray computed tomography. Scans were processed by a bespoke methodology that segmented vessels, facilitating diameter measurements. Though significant differences were not found comparing stem-average vessel section diameters among cultivars, they were found when comparing diameter distributions. Moreover, the measurements indicated that although vessel diameter distributions may play a role regarding the resistance of Leccino, it is unlikely they do for FS17. Considering Young–Laplace and Hagen–Poiseuille equations, we inferred differences in embolism susceptibility and hydraulic conductivity of the vasculature. Our results suggest susceptible cultivars, having a greater proportion of larger vessels, are more vulnerable to air embolisms. In addition, results suggest that under certain pressure conditions, functional vasculature in susceptible cultivars could be subject to greater stresses than in resistant cultivars. These results support investigation into xylem morphological screening to help inform olive replanting. Furthermore, our framework could test the relevance of xylem geometry to disease resistance in other crops.

We present a genome assembly from an individual male Carcina quercana (the long-horned flat-body; Arthropoda; Insecta; Lepidoptera; Depressariidae). The genome sequence is 409 megabases in span. Most of the assembly (99.96%) is scaffolded into 30 chromosomal pseudomolecules, including the assembled Z sex chromosome. The complete mitochondrial genome was also assembled and is 15.3 kilobases in length. Gene annotation of this assembly on Ensembl identified 18,108 protein coding genes.

Antimicrobial resistance (AMR) is a threat to global public health. However, unsatisfactory approaches to directly measuring the AMR burden carried by individuals has hampered efforts to assess interventions aimed at reducing selection for AMR. Metagenomics can provide accurate detection and quantification of AMR genes within an individual person’s faecal flora (their gut “resistome”). Using this approach, we aimed to test the hypothesis that differences in antimicrobial use across different hospitals in the United Kingdom will result in observable differences in the resistome of individual patients. Three National Health Service acute Hospital Trusts with markedly different antibiotic use and Clostridioides difficile infection rates collected faecal samples from anonymous patients which were discarded after C. difficile testing over a period of 9 to 15 months. Metagenomic DNA was extracted from these samples and sequenced using an Illumina NovaSeq 6000 platform. The resulting sequencing reads were analysed for taxonomic composition and for the presence of AMR genes. Among 683 faecal metagenomes we found huge variation between individuals in terms of taxonomic diversity (Shannon Index range 0.10–3.99) and carriage of AMR genes (Median 1.50 genes/cell/sample overall). We found no statistically significant differences in diversity (median Shannon index 2.16 (IQR 1.71–2.56), 2.15 (IQR 1.62–2.50) and 2.26 (IQR 1.55–2.51)) or carriage of AMR genes (median 1.37 genes/cell/sample (IQR 0.70–3.24), 1.70 (IQR 0.70–4.52) and 1.43 (IQR 0.55–3.71)) at the three trusts respectively. This was also the case across the sample collection period within the trusts. While we have not demonstrated differences over place or time using metagenomic sequencing of faecal discards, other sampling frameworks may be more suitable to determine whether organisational level differences in antibiotic use are associated with individual-level differences in burden of AMR carriage.

Air travel mediates transboundary movement of SARS-CoV-2. To prepare for future pandemics, we sought to understand air passenger behaviour and perceived risk during the COVID-19 pandemic. This study of UK adults (n = 2103) quantified knowledge of COVID-19 symptoms, perceived health risk of contracting COVID-19, likelihood of returning to the UK with COVID-19 symptoms, likelihood to obey self-quarantining guidelines, how safe air travellers felt when flying during the pandemic (n = 305), and perceptions towards face covering effectiveness.Overall knowledge of COVID-19 symptoms was poor. Men and younger age groups (18–44) were less informed than women and older age groups (44 +). A significant proportion (21%) of the population would likely travel back to the UK whilst displaying COVID-19 symptoms with many expressing that they would not fully comply with self-isolation guidelines. Overall, males and younger age groups had a reduced perceived personal risk from contracting COVID-19, posing a higher risk of transporting SARS-CoV-2 back to the UK. Poor passenger knowledge and behaviour undermines government guidelines and policies aimed at preventing SARS-CoV-2 entry into the UK. This supports the need for stricter, clearer and more targeted guidelines with point-of-departure viral testing and stricter quarantining upon arrival.

Multiple fungal species, including potential opportunistic pathogens have been previously identified in water systems. Here, we investigated over 250 restroom sink fungal communities across a university campus and evaluated their diversity and core taxa present. Remarkable similarity in mycobial community composition was observed across buildings with Ascomycota consistently dominating. We found a core mycobiome independent of the building sampled, that included Exophiala species, potential opportunistic pathogenic black yeasts. Other prevalent and dominant taxa included Saccharomyces and Fusarium, common built environment fungi. The frequent presence of Malassezia, a common skin commensal, showed the external influence of human activities as a source of fungi to sinks. The study represents a novel exploration of sink P-traps mycobial communities from a public area and highlights their importance as reservoirs of possible pathogenic fungi, as well as emphasizing the relevance of further research in this understudied ecosystem within the built environment.

Increasing interest in the use of microbial metrics to evaluate soil health raises the issue of how fine-scale heterogeneity can affect microbial community measurements. Here we analyse bacterial and fungal communities of over 100 soil samples across 17 pasture farms and evaluate beta diversity at different scales. We find large variation in microbial communities between different points in the same field, and if Aitchison distance is used we find that within-field variation is as high as between-farm variation. However, if Bray-Curtis or Jaccard distance are used this variation is partially explained by differences in soil pH and vegetation and is higher under mob grazing for fungi. Hence, field scale variation in microbial communities can impact the evaluation of soil health.

Apex predators are good indicators of environmental pollution since they are relatively long-lived and their high trophic position and spatiotemporal exposure to chemicals provides insights into the persistent, bioaccumulative and toxic (PBT) properties of chemicals. Although monitoring data from apex predators can considerably support chemicals’ management, there is a lack of pan-European studies, and longer-term monitoring of chemicals in organisms from higher trophic levels. The present study investigated the occurrence of contaminants of emerging concern (CECs) in 67 freshwater, marine and terrestrial apex predators and in freshwater and marine prey, gathered from four European countries. Generic sample preparation protocols for the extraction of CECs with a broad range of physicochemical properties and the purification of the extracts were used. The analysis was performed utilizing liquid (LC) chromatography coupled to high resolution mass spectrometry (HRMS), while the acquired chromatograms were screened for the presence of more than 2,200 CECs through wide-scope target analysis. In total, 145 CECs were determined in the apex predator and their prey samples belonging in different categories, such as pharmaceuticals, plant protection products, per- and polyfluoroalkyl substances, their metabolites and transformation products. Higher concentration levels were measured in predators compared to prey, suggesting that biomagnification of chemicals through the food chain occurs. The compounds were prioritized for further regulatory risk assessment based on their frequency of detection and their concentration levels. The majority of the prioritized CECs were lipophilic, although the presence of more polar contaminants should not be neglected. This indicates that holistic analytical approaches are required to fully characterize the chemical universe of biota samples. Therefore, the present survey is an attempt to systematically investigate the presence of thousands of

We use a national citizen science monitoring scheme to quantify how agricultural intensification affects honeybee diet breadth (number of plant species). To do this we used DNA metabarcoding to identify the plants present in 527 honey samples collected in 2019 across Great Britain. The species richness of forage plants was negatively correlated with arable cropping area, although this was only found early in the year when the abundance of flowering plants was more limited. Within intensively farmed areas, honeybee diets were dominated by Brassica crops (including oilseed rape). We demonstrate how the structure and complexity of honeybee foraging relationships with plants is negatively affected by the area of arable crops surrounding hives. Using information collected from the beekeepers on the incidence of an economically damaging bee disease (Deformed Wing Virus) we found that the occurrence of this disease increased where bees foraged in agricultural land where there was a high use of foliar insecticides. Understanding impacts of land use on resource availability is fundamental to assessing long-term viability of pollinator populations. These findings highlight the importance of supporting temporally timed resources as mitigation strategies to support wider pollinator population viability.

We present a genome assembly from an individual male Biston betularia (the peppered moth; Arthropoda; Insecta; Lepidoptera; Geometridae). The genome sequence is 405 megabases in span. The majority of the assembly (99.99%) is scaffolded into 31 chromosomal pseudomolecules, with the Z sex chromosome assembled.Gene annotation of this assembly on Ensembl has identified 12,251 protein coding genes.