The recent reinforcement of CoV surveillance in animals fuelled by the COVID-19 pandemic provided increasing evidence that mammals other than bats might hide further diversity and play critical roles in human infectious diseases. This work describes the results of a two-year survey carried out in Italy with the double objective of uncovering CoV diversity associated with wildlife and of excluding the establishment of a reservoir for SARS-CoV-2 in particularly susceptible or exposed species. The survey targeted hosts from five different orders and was harmonised across the country in terms of sample size, target tissues, and molecular test. Results showed the circulation of 8 CoV species in 13 hosts out of the 42 screened. Coronaviruses were either typical of the host species/genus or normally associated with their domestic counterpart. Two novel viruses likely belonging to a novel CoV genus were found in mustelids. All samples were negative for SARS-CoV-2, with minimum detectable prevalence ranging between 0.49% and 4.78% in the 13 species reaching our threshold sample size of 59 individuals. Considering that within-species transmission in white-tailed deer resulted in raising the prevalence from 5% to 81% within a few months, this result would exclude a sustained cycle after spillback in the tested species.

The mitotic-inhibiting herbicide pronamide controls susceptible annual bluegrass (Poa annua L.) pre- and postemergence, but in some resistant populations, postemergence activity is compromised, hypothetically due to a target-site mutation, lack of root uptake, or an unknown resistance mechanism. Three suspected pronamide-resistant (LH-R, SC-R, and SL-R) and two pronamide-susceptible (BS-S and HH-S) populations were collected from Mississippi golf courses. Dose–response experiments were conducted to confirm and quantify pronamide resistance, as well as resistance to flazasulfuron and simazine. Target sites known to confer resistance to mitotic-inhibiting herbicides were sequenced, as were target sites for herbicides inhibiting acetolactate synthase (ALS) and photosystem II (PSII). Pronamide absorption and translocation were investigated following foliar and soil applications. Dose–response experiments confirmed pronamide resistance of LH-R, SC-R, and SL-R populations, as well as instances of multiple resistance to ALS- and PSII-inhibiting herbicides. Sequencing of the α-tubulin gene confirmed the presence of a mutation that substituted isoleucine for threonine at position 239 (Thr-239-Ile) in LH-R, SC-R, SL-R, and BS-S populations. Foliar application experiments failed to identify differences in pronamide absorption and translocation between the five populations, regardless of harvest time. All populations had limited basipetal translocation—only 3% to 13% of the absorbed pronamide—across harvest times. Soil application experiments revealed that pronamide translocation was similar between SC-R, SL-R, and both susceptible populations across harvest times. The LH-R population translocated less soil-applied pronamide than susceptible populations at 24, 72, and 168 h after treatment, suggesting that reduced acropetal translocation may contribute to pronamide resistance. This study reports three new pronamide-resistant populations, two of which are resistant to two modes of action (MOAs), and one of which is resistant to three MOAs. Results suggest that both target site– and translocation-based mechanisms may be associated with pronamide resistance. Further research is needed to confirm the link between pronamide resistance and the Thr-239-Ile mutation of the α-tubulin gene.

Method development is at the heart of design research as methods are a formalised way to express knowledge about how aspects of design could or should be done. However, assuring that methods are in fact used in industry has remained a challenge. Industry will only use methods that they can understand and that they feel will give them benefit reliably. To understand the challenges involved in adopting a method, the method needs to be seen in context: it does not exist in isolation but forms a part of an ecosystem of methods for tackling related design problems. A method depends on the knowledge and skills of the practitioners using it: while a description of a method is an artefact that is a formalisation of engineering knowledge, a method in use constitutes a socio-technical system depending on the interaction of human participants with each other as well as with the description of the method, representations of design information and, often, tools for carrying out the method’s tasks. This paper argues that crucial factors in the adoption of methods include how well they are described and how convincingly they are evaluated. The description of a method should cover its core idea, the representations in which design information is described, the procedure to be followed, its intended use, and the tools it uses. The account of a method’s intended use should cover its purpose, the situations or product types within its scope, its coverage of kinds of problems within its scope, its expected benefit and conditions for its use. The different elements need to be evaluated separately as well as the method as an integrated whole. While verification and validation are important for some elements of methods, it is rarely possible to prove the validity of a method. Rather the developers of methods need to gather sufficient evidence that a method will work within a clearly articulated scope. Most design methods do not have binary success criteria, and their usefulness in practice depends as much on simplicity and usability as on the outcomes they produce. Evaluation should focus on how well they work, and how they can be customised and improved.

Nothofagus alessandrii, categorized as Endangered on the IUCN Red List, is an endemic, deciduous tree species of the coastal range of central Chile. We assessed the effects of fire severity, invasion by the exotic fire-prone Pinus radiata, and land-cover composition and configuration of the landscape on the resilience of fragments of N. alessandrii after a mega-fire in 2017. We used remote sensing data to estimate land-use classes and cover, fire severity and invasion cover of P. radiata. We monitored forest composition and structure and post-fire responses of N. alessandrii forests in situ for 2 years after the mega-fire. In the coastal Maule region wildfires have been favoured by intense drought and widespread exotic pine plantations, increasing the ability of fire-adapted invasive species to colonize native forest remnants. Over 85% of N. alessandrii forests were moderately or severely burnt. The propagation and severity of fire was probably amplified by the exotic pines located along the edges of, or inside, the N. alessandrii fragments and the highly flammable pine plantations surrounding these fragments (> 60% of land use is pine plantations). Pinus radiata, a fire-adapted pioneer species, showed strong post-fire recruitment within the N. alessandrii fragments, especially those severely burnt. Positive feedback between climate change (i.e. droughts and heat waves), wildfires and pine invasions is driving N. alessandrii forests into an undesirable and probably irreversible state (i.e. a landscape trap). A large-scale restoration programme to design a diverse and less flammable landscape is needed to avoid the loss of these highly threatened forest ecosystems.

Tree-ring chronologies encode interannual variability in forest growth rates over long time periods from decades to centuries or even millennia. However, each chronology is a highly localized measurement describing conditions at specific sites where wood samples have been collected. The question whether these local growth variabilites are representative for large geographical regions remains an open issue. To overcome the limitations of interpreting a sparse network of sites, we propose an upscaling approach for annual tree-ring indices that approximate forest growth variability and compute gridded data products that generalize the available information for multiple tree genera. Using regression approaches from machine learning, we predict tree-ring indices in space and time based on climate variables, but considering also species range maps as constraints for the upscaling. We compare various prediction strategies in cross-validation experiments to identify the best performing setup. Our estimated maps of tree-ring indices are the first data products that provide a dense view on forest growth variability at the continental level with 0.5° and 0.0083° spatial resolution covering the years 1902–2013. Furthermore, we find that different genera show very variable spatial patterns of anomalies. We have selected Europe as study region and focused on the six most prominent tree genera, but our approach is very generic and can easily be applied elsewhere. Overall, the study shows perspectives but also limitations for reconstructing spatiotemporal dynamics of complex biological processes. The data products are available at https://www.doi.org/10.17871/BACI.248.

The transient pressure field around a moderately thick airfoil is studied as it undergoes ramp-type pitching motions at high Reynolds numbers and low Mach numbers. A unique set of laboratory experiments were performed in a high-pressure wind tunnel to investigate dynamic stall at chord Reynolds numbers in the range of $0.5\times 10^6\leq Re _c\leq 5.5\times 10^6$ in the absence of compressibility effects. In addition to variations of mean angle and amplitude, pitching manoeuvres at reduced frequencies in the range of $0.01\leq k\leq 0.40$ were studied by means of surface-pressure measurements. Independently of the parameter variations, all test cases exhibit a nearly identical stall behaviour characterized by a gradual trailing-edge stall, in which the dynamic stall vortex forms approximately at mid-chord. The location of the pitching window with respect to the Reynolds-number-dependent static stall angle is found to define the temporal development of the stall process. The time until stall onset is characterized by a power law, where a small excess of the static stall angle results in a drastically prolonged stall delay. The reduced frequency exhibits a decrease in impact on the stall development in the case of angle-limited pitching manoeuvres. Beyond a critical reduced frequency, both load magnitudes and vortex evolution become reduced frequency independent and instead depend on the geometry of the motion and the convective time scale, respectively. Overall, the characteristics of vortex evolution induced by dynamic stall show remarkable similarities to the framework of optimal vortex formation reported in Gharib et al. (J. Fluid Mech., vol. 360, 1998, pp. 121–140). The data from this study are publicly available at https://doi.org/10.34770/b3vq-sw14.

The study followed 6-year-old children in Canadian French Immersion for three years to investigate the effect of home language background on acquisition of French, the language of schooling. None of the children knew French before beginning the program. French proficiency was indicated by French vocabulary and verbal fluency tasks. A language background questionnaire was used to (a) assign children to monolingual or bilingual groups and (b) provide a continuous score for degree of bilingual experience. Categorical analyses showed bilingual children had smaller English vocabulary than monolingual children when they entered the program. For French vocabulary, categorical comparisons revealed no language group differences in the first two years but higher French scores for bilingual children in the third year. In contrast, analyses of the continuous scores revealed a relation between more bilingual experience and higher French vocabulary throughout. Similarly, categorical analyses of verbal fluency results indicated no significant language group differences for either semantic or phonological fluency, but continuous analyses of semantic fluency showed an association between more bilingual experience and better outcomes in each year. These results suggest that language experience impacts progress in learning the language of schooling and that different analytic approaches reveal different aspects of the pattern.

The control of human visceral leishmaniasis (VL) is hard since there are no vaccines available as well as the treatment is hampered by toxicity and resistant parasites. Furthermore, as human, and canine VL causes immunosuppression, the combination of drugs with immunostimulatory agents is interesting to upregulate the immunity, reducing side-effects, improving treatment approaches against disease. Herein, we assessed the immunochemotherapy using miltefosine along with a vaccine formulated by Leishmania braziliensis antigens + saponin + monophosphoryl lipid-A (LBSapMPL) in L. infantum-infected hamsters. Two months after infection, the animals received treatments, and after 15 days they were evaluated for the treatment effect. The potential anti-Leishmania effect of miltefosine + LBSapMPL-vaccine was revealed by a specific immune response activation reflecting in control of spleen parasitism using half the miltefosine treatment time. The treated animals also showed an increase of total and T-CD4 splenocytes producing IFN-γ and TNF-α and a decrease of interleukin-10 and anti-Leishmania circulating IgG. In addition, it was demonstrated that the control of spleen parasitism is related to the generation of a protective Th1 immune response. Hence, due to the combinatorial action of miltefosine with LBSapMPL-vaccine in immunostimulating and controlling parasitism, this immunochemotherapy protocol can be an important alternative option against canine and human VL.

Lack of knowledge about iodine has been suggested as a risk factor for iodine deficiency in pregnant women, but no studies have addressed this issue in Portugal. So, the aim of this study was to investigate iodine knowledge among Portuguese pregnant women and its association with iodine status. IoMum, a prospective observational study, included 485 pregnant women recruited at Centro Hospitalar e Universitário de S. João, Porto, between the 10th and 13th gestational weeks. Partial scores for knowledge on iodine importance, on iodine food sources or on iodised salt were obtained through the application of a structured questionnaire. Then, a total iodine knowledge score was calculated and grouped into low, medium and high knowledge categories. Urinary iodine concentration (UIC) was measured in spot urine samples by inductively coupled plasma MS. Of the pregnant women, 54 % correctly recognised iodine as important to neurocognitive development, 32 % were unable to identify any iodine-rich food and 71 % presented lack of knowledge regarding iodised salt. Of the women, 61 % had a medium total score of iodine knowledge. Knowledge on iodine importance during pregnancy was positively associated with iodine supplementation and also with UIC. Nevertheless, median UIC in women who correctly recognised the importance of iodine was below the cut-off for adequacy in pregnancy (150 µg/l). In conclusion, knowledge on iodine importance is positively associated with iodine status. Despite this, recognising iodine importance during pregnancy may not be sufficient to ensure iodine adequacy. Literacy-promoting actions are urgently needed to improve iodine status in pregnancy.

The role of milk and dairy products in supplying iodine to pregnant women is unknown in Portugal. The aim of this study was to evaluate the association between milk and dairy product consumption and the iodine status of pregnant women in the IoMum cohort of the Oporto region. Pregnant women were recruited between 10 and 13 weeks of gestation, when they provided a spot urine sample and information on lifestyle and intake of iodine-rich foods. Urinary iodine concentration (UIC) was determined by inductively coupled plasma MS. A total of 468 pregnant women (269 iodine supplement users and 199 non-supplement users) were considered eligible for analysis. Milk (but not yogurt or cheese) intake was positively associated with UIC, in the whole population (P = 0·02) and in the non-supplement users (P = 0·002), but not in the supplement users (P = 0·29). In non-supplement users, adjusted multinomial logistic regression analysis showed that milk consumption <3 times/month was associated with a five times increased risk of having UIC < 50 µg/l when compared with milk consumption ≥2 times/d (OR 5·4; 95 % CI 1·55, 18·78; P = 0·008). The highest UIC was observed in supplement users who reported consuming milk once per d (160 µg/l). Milk, but not yogurt or cheese, was positively associated with iodine status of pregnant women. Despite the observed positive association, daily milk consumption may not be sufficient to ensure adequate iodine intake in this population.

Trematode prevalence and abundance in hosts are known to be affected by biotic drivers as well as by abiotic drivers. In this study, we used the unique salinity gradient found in the south-western Baltic Sea to: (i) investigate patterns of trematode infections in the first intermediate host, the periwinkle Littorina littorea and in the downstream host, the mussel Mytilus edulis, along a regional salinity gradient (from 13 to 22) and (ii) evaluate the effects of first intermediate host (periwinkle) density, host size and salinity on trematode infections in mussels. Two species dominated the trematode community, Renicola roscovita and Himasthla elongata. Salinity, mussel size and density of infected periwinkles were significantly correlated with R. roscovita, and salinity and density correlated with H. elongata abundance. These results suggest that salinity, first intermediate host density and host size play an important role in determining infection levels in mussels, with salinity being the main major driver. Under expected global change scenarios, the predicted freshening of the Baltic Sea might lead to reduced trematode transmission, which may be further enhanced by a potential decrease in periwinkle density and mussel size.