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    Towards a unified understanding of human–nature interactions

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    Mixoplankton interferences in dilution grazing experiments

    Our results show that Chl a alone is not an adequate proxy for prey growth rates in dilution grazing experiments when mixoplankton are present5,10. Chlorophyll is, in any case, a poor proxy for phototrophic plankton biomass31 because of inter-species variations, and also for the photoacclimation abilities of some species (for which very significant changes can occur within a few hours). The problem extends to the involvement of mixoplanktonic prey and grazers. Nevertheless, even very recent studies continue to rely on this parameter for quantifications of grazing despite acknowledging the dominance, both in biomass and abundance, of mixoplanktonic predators in their system30. Moreover, the detailed analysis of the species-specific dynamics revealed that different prey species are consumed at very different rates. In our experiments, and contrary to expectations (see32,33, and Fig. S1 in the Supplementary Information), C. weissflogii was only actively ingested in the ciliate experiment and, according to the results from the control bottles (Table 2), not by M. rubrum (see Fig. 4 and Fig. S1a).Certainly, it is not the first time that a negative selection against diatoms has been seen; for example, Burkill et al.34 noticed that diatoms were less grazed by protist grazers than other phytoplankton species, as assessed by a dilution technique paired with High-Performance Liquid Chromatography for pigment analysis. Using the same method, Suzuki et al.35 reported that diatoms became the dominant phytoplankton group, which suggests that other groups were preferentially fed upon. Calbet et al.36, in the Arctic, also found only occasional grazing over the local diatoms. In our study, diatoms were not only not consumed, but the presence of dinoflagellates appeared to contribute to their growth (Fig. 4), this relationship being partly dependent on the concentration of the predator (see Fig. 2c, d). This result could be a direct consequence of assimilation and use of compounds (e.g.,37,38) released by microplankton such as ammonium (e.g.,39,40) and urea (e.g.,41), which were not supplied in the growth medium, but which would have supported prey growth. Alternatively, this unexpected outcome may have been a consequence of the selective ingestion of R. salina by the two predators, relieving the competition for nutrients and light and resulting in a higher growth rate of the diatom in the presence of the predators. We cannot rule out the fact that diatoms sink faster than flagellates which, as the bottles were not mixed during most of the incubation period (although gently mixed at every sampling point), may have also involuntarily decreased ingestion rates on C. weissflogii. Still, one C. weissflogii cell contains, on average, ca. 2.5 times more Chl a than one R. salina cell (initial value excluded, see Table 3). Taken together with the preference for R. salina it is not surprising that the proportion of total Chl a represented by the diatoms increased over time, in particular in the L/D treatment (Figs. 6a, c and 7a, c).Table 3 Chl a content (pg Chl a cell−1) of the target species at each sampling point as calculated from the control bottles.Full size tableAnother factor clearly highlighted by our experiments, is that protozooplankton themselves contribute a significant portion of the total chlorophyll of the system (due to ingested Chl a), in particular at the beginning of the incubation (see Figs. 6 and 7); this being invariably ignored in a traditional dilution experiment. The high Chl a detected inside the protozooplanktonic grazers at the beginning of the incubations could suggest that the system was initially not in equilibrium, and that this was the result of superfluous feeding (e.g.,42). This would, nevertheless, be surprising since we required ca. 1 h to collect the initial samples (t = 0 h) after joining all the organisms together (see the section “Dilution grazing experiments” in the “Methods” section); previous studies, like the one on G. dominans and Oxyrrhis marina by Calbet et al.42, showed that the hunger response and consequent vacuole replenishment occurred in ca. 100 min for very high prey concentrations and it is expected to decrease at lower prey concentrations as the ones used in our study. Therefore, even if one assumes that the first 4 h of incubation are a result of superfluous feeding, after 24 h, the “estimated”, “observed”, and “from dilution slope” grazing estimates are not significantly different to those displayed in Fig. 5 (P  > 0.05 in all instances) and, therefore, we can assume that the hunger response was likely irrelevant (e.g.,43) and did not mask our results. In any case, as stated before, an actual field grazing dilution experiment also suffers from similar problems, because grazers and prey are suddenly diluted and not pre-adapted to distinct food concentrations. Nevertheless, this is not novel information, since Chl a and its degradation products have been found inside several protozooplankton species from different phylogenetic groups immediately after feeding44 and even after some days without food45. An increase in intracellular Chl a concentrations immediately after feeding has also been found in mixoplankton46,47, on which this increase is derived both from ingested prey as well as from new synthesis of their own Chl a. Additionally, several experiments with Live Fluorescently Labelled Algae (LFLA) show that predators (irrespective of their trophic mode) seem to maximise the concentration of intracellular prey shortly after the initiation of the incubation (e.g.,48; Ferreira et al., submitted). Indeed, some authors have even been able to measure photosynthesis in protozooplankton, like the ciliates Mesodinium pulex49 and Strombidinopsis sp.50.The fact that Chl a is a poor indicator of phytoplankton biomass and the inherent consequences discussed so far can be solved by the quantification of the prey community abundance (e.g.,51) by microscopy or by the use of signature pigments for each major phytoplankton group. The latter method, however, is not as thorough as the former, since rare are the cases where one pigment is exclusively associated with a single group of organisms (see52 and references therein). In any case, any pigment-based proxy is subject to the same problems, as identified by Kruskopf & Flynn31. Irrespective of the quantification method, it has been made evident that the different algae are consumed at different rates (e.g., pigments10,34,35; microscopy5,36).Prey selection in protistan grazers is a common feature (e.g.,23,26,27,28). Given the diversity of grazers in natural communities and the array of preferred prey that each particular species possesses, it is logical to think that dilution experiments will capture the net community response properly. Likewise, grazers interact with each other through toxins, competition, and intraguild predation among other factors. An example of intraguild predation could be the observed on K. armiger by G. dominans (see Figs. 2f and 4 and Table 1), which caused an average loss of ca. 18.72 pg of K. armiger carbon per G. dominans per hour in the D treatment. Interestingly, in the same treatment, a slight negative effect of K. armiger on its predator G. dominans can also be deduced (i.e., positive g, Table 1), resulting in an average loss of ca. 0.33 pg G. dominans carbon per K. armiger per hour. This could be a consequence of algal toxins, since K. armiger is a known producer of karmitoxin22, whose presence may have negative effects even on metazoan grazers21. Regarding ciliates, none of the species used is a known producer of toxic compounds, which suggests that the average loss of ca. 1.25 pg M. rubrum carbon per hour in the D treatment was due to S. arenicola predation. Altogether, it seems clear from our data that intraguild predation cannot be ignored when analysing dilution experiments (Fig. 4). Furthermore, our results clearly show that single functional responses cannot be used to extrapolate community grazing impacts, as evidenced by the differences in estimated and measured ingestion rates based on the disappearance of prey in combined grazers experiments (Fig. 5). Nevertheless, this is a relatively common procedure (e.g.,53 and references therein). Often in modelling approaches, individual predator’s functional responses have been used to extrapolate prey selectivity and community grazing responses27; in reality complex prey selectivity functions are required to satisfactorily describe prey selectivity and inter-prey allelopathic interactions54.It is, however, also evident that the measured ingestion rates in combined grazers experiments were not the same as those calculated from the slope of the dilution grazing experiment. This raises the question of why was that the case. It is well known that phytoplankton cultures, when extremely diluted, show a lag phase of different duration55 which has been attributed to the net leakage of metabolites56. Assuming that the duration of the lag phase will be dependent on the level of dilution, it seems reasonable to deduce that after ca. 24 h the instantaneous growth rates (µ) in the most diluted treatments will be lower than that of the undiluted treatments. This has consequences, not only for the estimated prey growth rates but also for the whole assessment of the grazing rate, due to the flattening of the regression line (i.e., the decrease in the computed growth rate). This artefact may be more evident in cultures acclimated to very particular conditions (as the laboratory cultures used in this study) than in nature.Another important finding of our research is the importance of light on the correct expression of the feeding activity by both mixoplankton and protozooplankton. We noticed that irrespective of the light conditions, all species exhibited a diurnal feeding rhythm (R. salina panels in Figs. 2 and 3), which is in accordance with earlier observations on protists (e.g.,29,57,58). The presence of light typically increased the ingestion rates. Additionally, the ingestion rates differed during the night period between L/D and D treatments, which implies that receiving light during the day is also vital in modulating the night behaviour of protoozoo- and mixoplankton. In particular, mixoplankton grazing is usually affected by light conditions, typically increasing (e.g.,32,59), but also sometimes decreasing(e.g.,60) in the presence of light. Different irradiance levels can also affect the magnitude of ingestion rates both in protozoo- and mixoplankton (see61 and references therein).For those reasons, we hoped for a rather consistent pattern among our protists that would help us discriminate mixoplankton in dilution grazing experiments. As a matter of fact, based on the results from Arias et al.29, we expected that in the dinoflagellate experiment, the D treatment would have inhibited only the grazing of K. armiger, enabling a simple discrimination between trophic modes. The reality did not meet the expectations since the day and night-time carbon-specific ingestion rates (as assessed using the control bottles, Table 2) of K. armiger were respectively higher and equal than those of G. dominans. Conversely, in the ciliate experiment, protozooplankton were the major grazers in our incubations regardless of the day period and light conditions. This response was not as straightforward as one would expect it to be because M. rubrum has been recently suggested to be a species complex containing at least 7 different species (62 and references therein), which hinders any possible conjecture on their grazing impact. Indeed, the uneven responses found between and within trophic modes precluded such optimistic hypothetical procedure.The D treatment in the present paper illustrated the importance of mimicking natural light conditions, a factor also addressed in the original description of the technique by Landry and Hassett1. It is crucial for the whole interpretation of the dilution technique that incubations should be conducted in similar light (and temperature) conditions as the natural ones to allow for the continued growth of the phototrophic prey. However, here we want to stress another aspect of the incubations: should they start during the day or the night? Considering our (and previous) results on diel feeding rhythms, and on the contribution of each species to the total Chl a pool, it is clear that different results will be obtained if the incubations are started during the day or the night. Besides, whether day or night, organisms are also likely to be in a very different physiological state (either growing or decreasing). Therefore, we recommend that dilution experiments conducted in the field should always be started at the same period of the day to enable comparisons (see also Anderson et al.14 for similar conclusions on bacterivory exerted by small flagellates). Ideally, incubations would be started at different times of the day to capture the intricacies of the community dynamics on a diel cycle. Nevertheless, should the segmented analysis be impossible, we argue that the right time to begin the incubations would be during the night, as this is the time where ingestion rates by protozooplankton are typically lower (e.g.,29,57,58, this study) and would, consequently, reduce their quota of Chl a in the system.Lastly, we want to stress that we are aware that our study does not represent natural biodiversity because our experiments were conducted in the laboratory with a few species. Nevertheless, we attempted to use common species of wide distribution for each major group of protists to provide a better institutionalisation of our conclusions. Further to the choice of predator and prey is their concentrations and proportions. Being a laboratory experiment designed to understand fundamental mechanisms within a dilution grazing experiment, we departed from near saturating food conditions from where we started the dilution series. In nature, the concentrations that we used may be high but are not unrealistic, and actually lower than in many bloom scenarios. We included diatoms at high concentrations, even knowing that they are not the preferred prey of most grazers34, because diatoms are very abundant in many natural ecosystems and to stress the point of food selection within the experiment. For sure, using different proportions of prey would have rendered different results. However, as previously mentioned, our aim was not to seek flaws in the dilution technique, but to understand the role of mixoplankton in these experiments and the complex trophic interactions that may occur within. Ultimately, with our choice of prey and their concentrations, we have proven that when there is no selection for a massively abundant prey, the use of Chl a as a proxy for community abundances may underestimate actual grazing rates.Some other aspects of our experiments may also be criticised because they do not fully match a standard dilution experiment. For instance, we manipulated light, adding complexity to the study. However, this manipulation enabled the deepening into the drivers of the mixoplanktonic and protozooplanktonic grazing responses. Another characteristic, perhaps awkward, of our study is that we allowed the grazers to deplete their prey before starting the experiment. One may argue this procedure does not mimic the natural previous trophic history a grazer may have in nature. Yet, in nature, when facing a dilution experiment, it is impossible to ascertain whether the organisms are encountering novel prey or not. Indeed, they (prey and predator) could have just migrated into such conditions, or be subject to famine, or just moved from a food patch. In any case, it is true that a consistent “hunger response” would have affected our initial grazing values, biasing grazing rate estimates. To overcome this artefact, we let the grazers feed for about one hour before starting the actual dilution assay (see the “Methods” section). From that point on, any dilution is, in fact, an abrupt alteration of the food scenario, which is likely more important than the previous trophic history of the grazer.In summary, with these laboratory experiments, we have presented evidence calling for a revision of the use of chlorophyll in dilution grazing experiments5,10, and we have highlighted the need to observe the organismal composition of both initial and final communities to better understand the dynamics during the dilution grazing experiments51. This approach will not incorporate mixoplanktonic activity into the dilution technique per se however if combined with LFLA (see5,17), a semi-quantitative approach to disentangle the contribution of mixoplankton to community grazing could be achieved (although not perfect). An alternative (and perhaps more elegant) solution could be the integration of the experimental technique with in silico modelling. The modelling approaches of the dilution technique have already been used, for example, to disentangle niche competition63 and to explore nonlinear grazer responses20. We believe that our experimental design and knowledge of the previously indicated data could be of use for the configuration of a dilution grazing model, which could then be validated in the field (and, optimistically, coupled to the ubiquitous application of the dilution technique across the globe). We cannot guarantee that having a properly constructed model that mimics the dilution technique will be the solution to the mixoplankton paradigm. However, it may provide a step towards that goal as it could finally shed much-needed light on the mixo- and heterotrophic contributions to the grazing pressure of a given system. To quote from the commentary of Flynn et al.6, it could provide the answer to the question of whether mixoplankton are de facto “another of the Emperor’s New Suit of Clothes” or, “on the other hand (…) collectively worthy of more detailed inclusion in models”. More

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    Estimating and predicting snakebite risk in the Terai region of Nepal through a high-resolution geospatial and One Health approach

    Our results showed that covariates at different geographical scales (national and local) may have important effects on the risk of snakebite, both for humans and animals. The results indicate that the risk of snakebite in the Terai varies at national scale between clusters and at local scale between households. The evaluation of the final models without spatial random components and the worsening of the models’ goodness of fit as a result highlighted how snakebite risk and its determining factors are indeed spatially structured.A strong association between high snakebite incidence and mortality, and poverty was established from the analysis of 138 countries affected by the disease32. In this study, we identified the PPI, an indicator for poverty, as a highly influential risk-increasing factor for humans. This not only confirms the critical role of poverty as a driver for this Neglected Tropical Disease, but also offers the possibility to use a standardized index at individual household scale for similar studies. Chaves et al.33 used the Poverty Gap, which is a simpler index expressing how far a person is from the average national poverty line, but to our knowledge, no study has used PPI for snakebite in any way. Applying PPI as a snakebite risk predictor also addresses previous expert calls for an Ecohealth approach to consider the relationship between the structural characteristics of houses, poverty, and snakebite34.Three of the survey covariates had significant effects on the odds of snakebite. Food storage and straw storage increased them, while sleeping on the floor reduce them. The effect of the first two covariates is likely to be related to prey availability, represented by rodents, which are attracted by food and shelter sources. Both food and straw are very often stored near dwellings, which in the end multiply the number of possible encounters between humans, domestic animals, and the hunting snakes20. The expected snakebite risk reduction effect by sleeping on the floor is more complex though. Previously, a higher snakebite incidence was reported among rural Hindus in Maharashtra, India, due to their custom of sleeping on the floor35,36, while in Nepal, Chappuis et al. did not find any protective effect or significant difference in snakebite cases between sleeping on a cot or on the floor37. This result, nevertheless, might be influenced again by regional customs that make sleeping on the floor more common in eastern Terai (71.1% of all affirmative answers to this question), and second, by the commonly acknowledged prevalence of kraits (Bungarus spp.) in western Terai, which are the species most commonly linked to bites to people sleeping on the floor while hunting at night inside houses22,38. This geographic separation, between the human behaviour and the distribution of the species considered to cause most bites linked to it, could explain the observed shift in the odds towards a reduction effect. This effect should be further explored in localized studies designed to capture behavioural differences in humans and snakes.For both the general human risk model and its equivalent prediction model, the covariate Distance to water had a significant risk-increasing effect. For each additional km in distance from permanent water sources, the odds of snakebite increased by 1.38 and 1.51 times, respectively. From a human perspective and in this socio-economic framework, it would be important to consider not only the distance to water, but also the path taken to get the water (or any other resource). If this path would lead a person through grasslands and open fields, this could imply an increased risk of snakebite. From an ecological perspective, there are two important aspects to consider in relation to water sources. One is, as in this study, the distance from large, constant water sources, which usually represent stable environments subject to less hydric stress. The second (not considered here) are the human-made water sources, such as ponds, reservoirs, and paddy fields that change often, are usually closer to human dwellings, and are known to attract some medically important venomous snakes (MIVS)5. Studies on snake migration and home range use have concluded that depending on species and ecological conditions, snakes can move between a few tens of meters per day and more than 10 km between seasons, while searching for water and prey resources38,39,40,41. In sub-tropical regions like the Terai, snakes living closer to continuous sources of water and vegetation should have easier access to a wider variety of prey. On the contrary, those living in agricultural areas might need to scout farther in the search for resources, encountering human-made waterbodies and prey, such as rodents42 and amphibians, abundant in this region10. Further studies considering all sources of water, and species ecology, biology and richness would be necessary to completely understand the effect of this and similar eco-physiological covariates.Another important factor was the NDVI, which is a commonly used value to express photosynthetic activity, leaf production and in summary the ‘greenness’ of the environment43. As is the case for other covariates, its interpretation depends on the study circumstances. In Iran, it was considered an indicator of prey availability for snakes and linked to snake habitat suitability14. Elevated NDVI values have been associated with higher number of hospitalizations in Nigeria and northern Ghana, in particular during the periods of high agricultural activity, which is also related to higher snake-human contact and higher snakebite incidence43. In our study, its ‘protective’ effect can indeed be the consequence of better access to prey associated with healthier ecosystems, explained in the Terai by the higher NDVI values of the multiple dense forests distributed along the region. In addition, the averaged NDVI values for agricultural areas should be lower than those for perennial forests, because they include the highs and lows of production and harvest.Environmental drivers like temperature and precipitation are common factors in geospatial analyses of snakebite13,14,17,44. They are found in many cases to be the main factors modulating the incidence or risk of snakebite, while varying in importance according to study conditions. For example, in Iran, precipitation seasonality was the most prevalent climatic covariate determining the habitat suitability leading to snakebite risk14, while in Mozambique, temperature seasonality was the predominant covariate13. Despite the Terai’s sub-tropical climate, the range of the average minimum temperature of the coldest month (BIO6) was 1.8–10.9 °C. For our snakebite risk analysis in animals, an increase of 10 °C of BIO6 between any two points represented an increase in the odds of snakebite of 23.41 times. For snakes, this range could be the difference between total lethargy and partial activity45, which could lead to increased numbers of snakebites. In addition, and according to the production and holding practices of domestic animals in the Terai, this temperature range can also represent the difference between animals (mainly ruminants) being kept in sheds when at the lower range limits, or being let out of them at the upper limits, which would again increase the chances of encounters with snakes.Similarly, for the animal model, pig density and sheep density, significantly influenced the variation in the risk of snakebite for animals in the Terai. This could be due to the conditions in which the animals and their feed are kept, favouring environments that are beneficial for either snakes or their prey. At more local scales, rather than the distribution, the presence of other animal species could instead be the factor associated with higher snakebite rates12. However, since the available data on domestic animal density was produced more than 10 years ago, and the animal population has grown substantially in the last years in Nepal, this outcome should be interpreted with caution.For the animal risk, the possession of an animal shed also significantly increased the odds of snakebite. Similar to straw storage, animal sheds and similar constructions offer some shelter and at the same time attract small (prey) animals, both of which are likely to attract snakes, increasing snakebite risk for the animals using the shed. If in addition, the sheds function as poultry coops, the snake hunting behaviour might be instead targeted towards chicks and chickens12. Mitigation measures such as raising the coop’s floor or securing openings with fine metal mesh have been suggested to reduce this risk12.The human modification of terrestrial systems was the only non-significant covariate in the animal risk model. However, as its strong, risk-reducing effect still seems to explain a lot of the response variation, it was retained. Its change in one unit, i.e., going from a pristine to fully modified environment, decreased the odds of snakebite by 0.13 (equivalent to 7.69 times), which agrees with previous national survey results from Sri Lanka21.For our human risk prediction model, four covariates were either significant or helped to explain the changes in the response. Distance to water and NDVI were clearly significant, and precipitation of the driest quarter (BIO17) and the mean annual temperature (BIO1) helped to explain some of the response variation with convincing, unambiguous effects. For BIO17, an increase of 100 mm of rain during the driest months of the year represented an odds-reduction effect equivalent to 8.33 times. This agrees with the results of distance to water, suggesting that the additional availability of resources during water shortage periods, i.e., almost four times more rain (BIO17 range: 18–71 mm), could locally improve ecological conditions for snakes also leading to less scouting and fewer human encounters. Previous studies have analysed the multilevel ecological effects of droughts, e.g., reducing snake prey and leading snakes to engage in riskier behaviours46,47. For BIO1, the protective effect was weaker. An increase of 10 °C represented a reduction of the odds of snakebite equivalent to 3.57 times. Average temperatures for specific locations are difficult to interpret, since they might depend on mild highs and lows, strong highs and lows, or relative combinations of both. Thus, despite having a relatively important effect on the response, this effect still might be the consequence of confounding and unknown interactions.Several other evaluated covariates, for both humans and animals, showed a negligible effect on describing the response, were not significant while having very large uncertainties, or both. Consequently, they were discarded as predicting factors. For the list of baseline covariates evaluated, see supplementary Table S1. For a complete list of available survey covariates, see Alcoba et al.27.Some of our discarded covariates have been important in other studies, for example, to quantify snakebite risk based on reclassification methods of covariates such as habitat suitability, species presence, or envenoming severity13,14,17,44,48. These methods are especially relevant when one species (or very few) is the cause of most snakebite cases, and has differentiated optimal and sub-optimal habitats. In Nepal, and particularly in the Terai, there are at least two, and sometimes more than 10 MIVS with overlapping distributions49. Thus, it could be said that practically the whole region offers suitable habitat for multiple MIVS. In addition, the impossibility of reliably identifying the species having bitten the surveyed victims hindered the use of single species in the analysis. In our analysis, species richness was removed, as it showed almost no effect on the response. A recent meta-analysis reported an equivalent result at global scale, finding no significant difference between the number of venomous snake species in tropical and temperate locations, while the number of snakebites is clearly higher in tropical areas50. These results suggested that high incidence of snakebite is unrelated to species richness, but instead related to other factors like the number of people working in agricultural environments21,32,50. Another important driver of snakebite incidence has been population density50. In our study, however, any possible effect from population density on the risk was diminished by the random selection of households at specific numbers during study design. This was later confirmed by the minimal effect of population density as covariate in the human risk analysis.This study presents a few limitations. For instance, despite the capacity of the INLA method to borrow strength from neighbouring observations and areas, the selection of adequate covariates with enough explanatory power still depends greatly on the number of snakebite cases, which even for a national scale study like this remains small. Also, some of the covariates with the strongest explanatory power came from our household survey, which prevented their use for generalized spatial prediction models. Concerning the animal risk analysis, due to the small number of snakebite cases we opted to aggregate all animal species and consider a grouped response. Thus, for a spatial analysis of animal risk, it was not worth it to consider each species, since that would dilute further an already sparse dataset in individual models and selection processes. Moreover, the data gathered for animals was dependent on the random selection of (human) households and unrelated to the current distribution of animal populations. This, in addition to the possible number of dry bites that go unnoticed, might be responsible for the low number of animal victims recorded (even combined across all species), making a more detailed analysis unfeasible.Despite the large number of covariates examined during our analysis, very few were useful to predict snakebite risk along the Terai. It is possible that confounders or other difficult-to-measure covariates could better explain the complex relationship between the ecology and biology of MIVS, socio-economic factors, human behavioural traits, and the circumstances around domestic animal keeping. This needs to be further explored, following a recent call for an overarching One Health and Ecohealth approach to better understand the drivers for snakebite risk, incidence, and mortality under specific situations34.In conclusion, snakebite is a multi-factorial disease and there is no possible universal approach to model its risk. Each model should be individually designed for each set of socio-economical, geographic, ecological, cultural, and environmental circumstances19. To better understand and address the snakebite problem, it is necessary to approach it, whenever possible, with local data collected at a national scale, so that the conclusions drawn can fuel appropriate national public health policies and actions. As long as people work, live, and keep their domestic animals in close contact with natural environments with MIVS, the risk of snakebite will be present. However, better understanding of the factors influencing that risk at the most granular scale possible, and the estimation of the populations at risk, can help to better target prevention and mitigation measures. For humans, this evidence can channel efforts towards improved access to treatment through the optimized stockpiling of antivenom, and the improvement, relocation or new construction of treating facilities, but more importantly, towards community education and sensitization in preventive campaigns51. Part of that preventive and educative efforts can be done at household level, by promoting and facilitating the use of protective equipment such as rubber boots, or the guidance on how to improve and adapt their immediate surroundings to make them ecologically less attractive for snakes and their prey. For domestic animals, this information could help better target awareness-raising activities for animal owners and implement mitigation strategies. For animals at higher risk, tailored interventions such as the improvement of housing conditions, regular cleaning of sheds and surrounding areas (e.g., from food waste and surrounding vegetation), and using light when animals are walked out of the enclosure at night could be deployed specifically as snakebite prevention measures52. It is also important to highlight that many of the factors analysed in this study affect most directly the snakes themselves, not only as snakebite agents, but also as a diverse group of species, differently affected by ecological, climatic and environmental factors in a multiplicity of settings shared with humans and domestic animals. It is therefore necessary to further investigate how those factors influence the behavioural and ecological traits of MIVS in order to truly understand this disease from a One Health viewpoint. At stake is the reduction of snakebite envenoming incidence rates in humans and animals, and of its possible long-term sequelae on human populations. More

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    Phage co-transport with hyphal-riding bacteria fuels bacterial invasion in a water-unsaturated microbial model system

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    Commensal Pseudomonas protect Arabidopsis thaliana from a coexisting pathogen via multiple lineage-dependent mechanisms

    Systemic co-infections of commensal Pseudomonas with an individual pathogenTo examine the ability of commensal Pseudomonas strains to protect host plants from members of a pathogenic Pseudomonas lineage, we made use of a local isolate collection [16]. We henceforth refer to an operational taxonomic unit (OTU) as reported in that study as “ATUE” (isolates from Around TUEbingen), and following previous findings [16, 17], we refer to the lineage ATUE5 as pathogenic, and to all non-ATUE5 lineages as commensals.We grew plants on MS agar and monitored plant growth and health by extracting the number of green pixels from images over time (illustration in Fig. 1A). Green pixel count and rosette fresh weight were strongly correlated (Supplementary Fig. S1; R2 = 0.92, p value  More

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    Carbon assimilating fungi from surface ocean to subseafloor revealed by coupled phylogenetic and stable isotope analysis

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