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Abstract: High-throughput sequencing (HTS) technologies and bioinformatic analyses are of growing interest to be used as a routine diagnostic tool in the field of plant viruses. The reliability of HTS workflows from sample preparation to data analysis and results interpretation for plant virus detection and identification must be evaluated (verified and validated) to approve this tool for diagnostics. Many different extraction methods, library preparation protocols, and sequence and bioinformatic pipelines are available for virus sequence detection. To assess the performance of plant virology diagnostic laboratories in using the HTS of ribosomal RNA depleted total RNA (ribodepleted totRNA) as a diagnostic tool, we carried out an interlaboratory comparison study in which eight participants were required to use the same samples, (RNA) extraction kit, ribosomal RNA depletion kit, and commercial sequencing provider, but also their own bioinformatics pipeline, for analysis. The accuracy of virus detection ranged from 65% to 100%. The false-positive detection rate was very low and was related to the misinterpretation of results as well as to possible cross-contaminations in the lab or sequencing provider. The bioinformatic pipeline used by each laboratory influenced the correct detection of the viruses of this study. The main difficulty was the detection of a novel virus as its sequence was not available in a publicly accessible database at the time. The raw data were reanalysed using Virtool to assess its ability for virus detection. All virus sequences were detected using Virtool in the different pools. This study revealed that the ribodepletion target enrichment for sample preparation is a reliable approach for the detection of plant viruses with different genomes. A significant level of virology expertise is needed to correctly interpret the results. It is also important to improve and complete the reference data.
Keywords: high-throughput sequencing, ribodepletion, interlaboratory comparison, test performance study, proficiency test, Virtool
Published in DiRROS: 19.07.2024; Views: 129; Downloads: 75
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Abstract: Research on marine microbial communities is growing, but studies are hard to compare because of variation in seawater sampling protocols. To help researchers in the inter-comparison of studies that use different seawater sampling methodologies, as well as to help them design future sampling campaigns, we developed the EuroMarine Open Science Exploration initiative (EMOSE). Within the EMOSE framework, we sampled thousands of liters of seawater from a single station in the NW Mediterranean Sea (Service d'Observation du Laboratoire Arago [SOLA], Banyuls-sur-Mer), during one single day. The resulting dataset includes multiple seawater processing approaches, encompassing different material-type kinds of filters (cartridge membrane and flat membrane), three different size fractionations (>0.22 µm, 0.22–3 µm, 3–20 µm and >20 µm), and a number of different seawater volumes ranging from 1 L up to 1000 L. We show that the volume of seawater that is filtered does not have a significant effect on prokaryotic and protist diversity, independently of the sequencing strategy. However, there was a clear difference in alpha and beta diversity between size fractions and between these and “whole water” (with no pre-fractionation). Overall, we recommend care when merging data from datasets that use filters of different pore size, but we consider that the type of filter and volume should not act as confounding variables for the tested sequencing strategies. To the best of our knowledge, this is the first time a publicly available dataset effectively allows for the clarification of the impact of marine microbiome methodological options across a wide range of protocols, including large-scale variations in sampled volume.
Keywords: marine microbiome, standardized sampling, inter-comparison, amplicon sequencing, microbial diversity, seawater sampling
Published in DiRROS: 12.07.2024; Views: 104; Downloads: 148
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Abstract: Efficient traffic management is essential in modern urban areas. The development of intelligent traffic flow prediction systems can help to reduce travel times and maximize road capacity utilization. However, accurately modeling complex spatiotemporal dependencies can be a difficult task, especially when real-time data collection is not possible. This study aims to tackle this challenge by proposing a solution that incorporates extensive feature engineering to combine historical traffic patterns with covariates such as weather data and public holidays. The proposed approach is assessed using a new real-world data set of traffic patterns collected in Ljubljana, Slovenia. The constructed models are evaluated for their accuracy and hyperparameter sensitivity, providing insights into their performance. By providing practical solutions for real-world scenarios, the proposed approach offers an effective means to improve traffic flow prediction without relying on real-time data.
Keywords: traffic modeling, time-series forecasting, traffic-count data set, machine learning, model comparison
Published in DiRROS: 28.09.2023; Views: 554; Downloads: 257
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Abstract: Tree heights are one of the most important aspects of forest mensuration, but data are often unavailable due to costly and time-consuming field measurements. Therefore, various types of models have been developed for the imputation of tree heights for unmeasured trees, with mixed-effects models being one of the most commonly applied approaches. The disadvantage here is the need of sufficient sample size per tree species for each plot, which is often not met, especially in mixed forests. To avoid this limitation, we used principal component analysis (PCA) for the grouping of similar plots based on the most relevant site descriptors. Next, we compared mixed-effects models with height-diameter models based on artificial neural networks (ANN). In terms of root mean square error (RMSE), mixed-effects models provided the most accurate tree height predictions at the plot level, especially for tree species with a smaller number of tree height measurements. When plots were grouped using the PCA and the number of observations per category increased, ANN predictions improved and became more accurate than those provided by mixed-effects models. The performance of ANN also increased when the competition index was included as an additional explanatory variable. Our results show that in the pursuit of the most accurate modelling approach for tree height predictions, ANN should be seriously considered, especially when the number of tree measurements and their distribution is sufficient.
Keywords: height-diameter models, national forest inventory, permanent sample plot, mixed forests, model comparison, principal component analysis
Published in DiRROS: 08.06.2022; Views: 686; Downloads: 314
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