Performance of T-Track® TB, a Novel Dual Marker RT-qPCR-Based Whole-Blood Test for Improved Detection of Active Tuberculosis.

Tuberculosis (TB) is one of the leading causes of death by an infectious disease. It remains a major health burden worldwide, in part due to misdiagnosis. Therefore, improved diagnostic tests allowing the faster and more reliable diagnosis of patients with active TB are urgently needed. This prospective study examined the performance of the new molecular whole-blood test T-Track® TB, which relies on the combined evaluation of IFNG and CXCL10 mRNA levels, and compared it to that of the QuantiFERON®-TB Gold Plus (QFT-Plus) enzyme-linked immunosorbent assay (ELISA). Diagnostic accuracy and agreement analyses were conducted on the whole blood of 181 active TB patients and 163 non-TB controls. T-Track® TB presented sensitivity of 94.9% and specificity of 93.8% for the detection of active TB vs. non-TB controls. In comparison, the QFT-Plus ELISA showed sensitivity of 84.3%. The sensitivity of T-Track® TB was significantly higher (p < 0.001) than that of QFT-Plus. The overall agreement of T-Track® TB with QFT-Plus to diagnose active TB was 87.9%. Out of 21 samples with discordant results, 19 were correctly classified by T-Track® TB while misclassified by QFT-Plus (T-Track® TB-positive/QFT-Plus-negative), and two samples were misclassified by T-Track® TB while correctly classified by QFT-Plus (T-Track® TB-negative/QFT-Plus-positive). Our results demonstrate the excellent performance of the T-Track® TB molecular assay and its suitability to accurately detect TB infection and discriminate active TB patients from non-infected controls.

A dataset of road-killed vertebrates collected via citizen science from 2014-2020.

Data on road-killed animals is essential for assessing the impact of roads on biodiversity. In most European countries data on road-killed huntable wildlife exists, but data on other vertebrate species (e.g. amphibians, reptiles, small mammals) is scarce. Therefore, we conducted a citizen science project on road-killed vertebrates as a useful supplement to data on huntable wildlife collected by public authorities. The dataset contains 15198 reports with 17163 individual road-killed vertebrates collected by 912 participants. The reports were made in 44 countries, but the majority of data was reported in Austria. We implemented a data validation routine which led to three quality levels. Reports in quality level 1 are published via GBIF, reports in quality level 2 via Zenodo and reports in quality level 3 were deleted. The dataset is relevant for the scientific community studying impacts of roads on fauna as well as for those who are responsible for road planning and implementing mitigation measures.

A decrease in reports on road-killed animals based on citizen science during COVID-19 lockdown.

BACKGROUND: To avoid the uncontrolled spread of COVID-19 in early 2020, many countries have implemented strict lockdown measures for several weeks. In Austria, the lockdown in early spring has led to a significant drop in human outdoor activities, especially in road traffic. In Project Roadkill, a citizen science project which aims to collect data on road-killed animals, we observed a significant decrease in reported roadkills. METHODS: By asking the citizen scientists through a survey how their travelling routines were affected, we investigated if the observed decrease in roadkills was grounded in less animals being killed by traffic, or in citizen scientists staying at home and thus reporting less road-killed animals. RESULTS: A majority of the respondents stated that they felt to have reported less roadkills during the lockdown, regardless if they changed their travelling routine or not. This observation in combination with the overall decrease in road traffic indicates that fewer animals were killed during the lockdown. We conclude that when analyzing citizen science data, the effects of lockdown measures on reporting behaviour should be considered, because they can significantly affect data and interpretation of these data.

Predicting spring migration of two European amphibian species with plant phenology using citizen science data.

Habitat fragmentation is one of the drivers for amphibian population declines globally. Especially in industrialized countries roads disrupt the seasonal migration of amphibians between hibernation and reproduction sites, often ending in roadkills. Thus, a timely installing of temporary mitigation measures is important for amphibian conservation. We wanted to find out if plant phenology can be a proxy in advance to determine the start of amphibian migration, since both phenomena are triggered by temperature. We analysed data of 3751 amphibian and 7818 plant phenology observations from citizen science projects in Austria between 2000 and 2018. Using robust regression modelling we compared the migration of common toads (Bufo bufo) and common frogs (Rana temporaria) with the phenology of five tree, one shrub, and one herb species. Results showed close associations between the migration of common frogs and phenological phases of European larch, goat willow and apricot. Models based on goat willow predict migration of common frog to occur 21 days after flowering, when flowering was observed on 60th day of year; apricot based models predict migration to occur 1 day after flowering, observed on the 75th day of year. Common toads showed weaker associations with plant phenology than common frogs. Our findings suggest that plant phenology can be used to determine the onset of temporary mitigation measures for certain amphibian species to prevent roadkills.

Contours of citizen science: a vignette study.

Citizen science has expanded rapidly over the past decades. Yet, defining citizen science and its boundaries remained a challenge, and this is reflected in the literature-for example in the proliferation of typologies and definitions. There is a need for identifying areas of agreement and disagreement within the citizen science practitioners community on what should be considered as citizen science activity. This paper describes the development and results of a survey that examined this issue, through the use of vignettes-short case descriptions that describe an activity, while asking the respondents to rate the activity on a scale from 'not citizen science' (0%) to 'citizen science' (100%). The survey included 50 vignettes, of which five were developed as clear cases of not-citizen science activities, five as widely accepted citizen science activities and the others addressing 10 factors and 61 sub-factors that can lead to controversy about an activity. The survey has attracted 333 respondents, who provided over 5100 ratings. The analysis demonstrates the plurality of understanding of what citizen science is and calls for an open understanding of what activities are included in the field.

Experimental indications of gardeners' anecdotes that snails interfere with invasive slugs.

The invasive Spanish slug (Arion vulgaris) is an important pest species in agriculture and horticulture in Europe. In the last decades it has spread across the continent where it outcompetes native slug and snail species, thus posing a threat for biodiversity. A popular anecdote suggests to promote Roman snails (Helix pomatia) in gardens because they are able to control A. vulgaris. We examined a potential interrelationship between these two species using a mesocosm experiment with lettuce plants. 13C-15N stable isotope labelling of lettuce allowed us to investigate interactions between Helix and Arion on weight gain/loss and herbivory. Additionally, we wanted to know whether different watering regimes (daily vs. every 3rd day watering of weekly amount) and earthworms alter these interactions. Egg predation of Helix on Arion eggs was further tested in a food-choice experiment. Arion showed a five times higher herbivory per body mass than Helix in a single-species setting. However, in mesocosms containing both species percentage of herbivory per body mass was significantly lower than in Arion-only mesocosms, especially when watered every three days. Overall isotope uptake via eaten lettuce was unaffected by the presence of the other species. Only very little predation (three out of 200 eggs) of Helix on Arion eggs was observed. Our results provide no evidence for a clear dismissal or confirmation of the popular gardener's anecdote that Helix snails have a negative effect on Arion abundance or herbivory.

Foliar Roundup application has minor effects on the compositional and functional diversity of soil microorganisms in a short-term greenhouse experiment.

The herbicide Roundup (and glyphosate, its active ingredient) is extensively used for weed control on a worldwide scale. It is absorbed after foliar application and quickly translocated inside the plant. In this study, we investigated the effects of Roundup speed, a commercial glyphosate formulation, on the structural composition (dominance of microbial groups, phospholipid fatty acid analysis - PLFA) and functional diversity (use of carbon sources, Multiple Substrate Induced Respiration - MSIR) of soil microorganisms. We specifically aimed at understanding the potential impact of biotic interactions on herbicide effects and included plants, earthworms, and endomycorrhizal fungi in the experimental setup. For this, we grew clover (Trifolium repens) in the greenhouse and added mycorrhizal inoculum (Glomus mosseae) and earthworms (Lumbricus terrestris) to the pots. Two weeks after foliar Roundup application and subsequent plant death, the pots were destructively sampled. The application resulted in a significant increase of microbial respiration (SIR) by approximately 30%. A multivariate analysis of the MSIR data exhibited small but significant differences between the microbial communities of treated and untreated pots, while no significant difference was apparent for the PLFA data. Bacterial PLFAs generally decreased following herbicide application, while mycorrhizal and fungal PLFAs were not affected. We did not find a consistent difference between the fatty acid markers of gram negative and gram positive bacteria. For all investigated parameters, there were highly significant differences between the upper (0-5 cm depth) and lower (5-10 cm) soil layers. The fact that rooting density differed by a factor of 3.5 between the two layers indicated that herbicide effects were especially pronounced in the clover rhizosphere and were likely due to changes in root exudate composition. We found significant, though very small, interactions between Roundup and other experimental factors (especially mycorrhizal inoculum).

Fox sightings in a city are related to certain land use classes and sociodemographics: results from a citizen science project.

BACKGROUND: Red foxes (Vulpes vulpes L.) have become successful inhabitants of urban areas in recent years. However, our knowledge about the occurrence, distribution and association with land uses of these urban foxes is poor, partly because many favoured habitats are on private properties and therefore hardly accessible to scientists. We assumed that citizen science, i.e. the involvement of the public, could enable researchers to bridge this information gap. We analysed 1179 fox sightings in the city of Vienna, Austria reported via citizen science projects to examine relationships between foxes and the surrounding land use classes as well as sociodemographic parameters. RESULTS: Conditional probabilities of encountering foxes were substantially higher in gardens, areas with a low building density, parks or squares as compared to agricultural areas, industrial areas or forests. Generalized linear model analyses showed that sociodemographic parameters such as education levels, district area, population density and average household income additionally improved the predictability of fox sightings. CONCLUSIONS: Reports of fox sightings by citizen scientists might help to support the establishment of wildlife management in cities. Additionally, these data could be used to address public health issues in relation with red foxes as they can carry zoonoses that are also dangerous to humans.

Amphibian and reptile road-kills on tertiary roads in relation to landscape structure: using a citizen science approach with open-access land cover data.

BACKGROUND: Amphibians and reptiles are among the most endangered vertebrate species worldwide. However, little is known how they are affected by road-kills on tertiary roads and whether the surrounding landscape structure can explain road-kill patterns. The aim of our study was to examine the applicability of open-access remote sensing data for a large-scale citizen science approach to describe spatial patterns of road-killed amphibians and reptiles on tertiary roads. Using a citizen science app we monitored road-kills of amphibians and reptiles along 97.5 km of tertiary roads covering agricultural, municipal and interurban roads as well as cycling paths in eastern Austria over two seasons. Surrounding landscape was assessed using open access land cover classes for the region (Coordination of Information on the Environment, CORINE). Hotspot analysis was performed using kernel density estimation (KDE+). Relations between land cover classes and amphibian and reptile road-kills were analysed with conditional probabilities and general linear models (GLM). We also estimated the potential cost-efficiency of a large scale citizen science monitoring project. RESULTS: We recorded 180 amphibian and 72 reptile road-kills comprising eight species mainly occurring on agricultural roads. KDE+ analyses revealed a significant clustering of road-killed amphibians and reptiles, which is an important information for authorities aiming to mitigate road-kills. Overall, hotspots of amphibian and reptile road-kills were next to the land cover classes arable land, suburban areas and vineyards. Conditional probabilities and GLMs identified road-kills especially next to preferred habitats of green toad, common toad and grass snake, the most often found road-killed species. A citizen science approach appeared to be more cost-efficient than monitoring by professional researchers only when more than 400 km of road are monitored. CONCLUSIONS: Our findings showed that freely available remote sensing data in combination with a citizen science approach would be a cost-efficient method aiming to identify and monitor road-kill hotspots of amphibians and reptiles on a larger scale.

Glyphosate herbicide affects belowground interactions between earthworms and symbiotic mycorrhizal fungi in a model ecosystem.

Herbicides containing glyphosate are widely used in agriculture and private gardens, however, surprisingly little is known on potential side effects on non-target soil organisms. In a greenhouse experiment with white clover we investigated, to what extent a globally-used glyphosate herbicide affects interactions between essential soil organisms such as earthworms and arbuscular mycorrhizal fungi (AMF). We found that herbicides significantly decreased root mycorrhization, soil AMF spore biomass, vesicles and propagules. Herbicide application and earthworms increased soil hyphal biomass and tended to reduce soil water infiltration after a simulated heavy rainfall. Herbicide application in interaction with AMF led to slightly heavier but less active earthworms. Leaching of glyphosate after a simulated rainfall was substantial and altered by earthworms and AMF. These sizeable changes provide impetus for more general attention to side-effects of glyphosate-based herbicides on key soil organisms and their associated ecosystem services.

Earthworm-mycorrhiza interactions can affect the diversity, structure and functioning of establishing model grassland communities.

Both earthworms and arbuscular mycorrhizal fungi (AMF) are important ecosystem engineers co-occurring in temperate grasslands. However, their combined impacts during grassland establishment are poorly understood and have never been studied. We used large mesocosms to study the effects of different functional groups of earthworms (i.e., vertically burrowing anecics vs. horizontally burrowing endogeics) and a mix of four AMF taxa on the establishment, diversity and productivity of plant communities after a simulated seed rain of 18 grassland species comprising grasses, non-leguminous forbs and legumes. Moreover, effects of earthworms and/or AMF on water infiltration and leaching of ammonium, nitrate and phosphate were determined after a simulated extreme rainfall event (40 l m(-2)). AMF colonisation of all three plant functional groups was altered by earthworms. Seedling emergence and diversity was reduced by anecic earthworms, however only when AMF were present. Plant density was decreased in AMF-free mesocosms when both anecic and endogeic earthworms were active; with AMF also anecics reduced plant density. Plant shoot and root biomass was only affected by earthworms in AMF-free mesocosms: shoot biomass increased due to the activity of either anecics or endogeics; root biomass increased only when anecics were active. Water infiltration increased when earthworms were present in the mesocosms but remained unaffected by AMF. Ammonium leaching was increased only when anecics or a mixed earthworm community was active but was unaffected by AMF; nitrate and phosphate leaching was neither affected by earthworms nor AMF. Ammonium leaching decreased with increasing plant density, nitrate leaching decreased with increasing plant diversity and density. In order to understand the underlying processes of these interactions further investigations possibly under field conditions using more diverse belowground communities are required. Nevertheless, this study demonstrates that belowground-aboveground linkages involving earthworms and AMF are important mediators of the diversity, structure and functioning of plant communities.