51. The International cookbook for wastewater practitioners : testing wastewater for public health, SARS-CoV-2Bernd Manfred Gawlik, Sara Comero, Daniel A. Deere, Gertjan Medema, Ismahane Remonnay, Jean-François Loret, Ana Maria de Roda Husman, Stephanie Rinck-Pfeiffer, Shelesh Agrawal, Ion Gutiérrez-Aguirre, 2024, final research report Abstract: During the 2020-2023 COVID-19 Global Public Health Emergency of International Concern, rapid progress was made with the application of Wastewater-Based Surveillance (WBS) (also known as Wastewater-Based Epidemiology (WBE)) to provide population-scale evidence of SARS-CoV-2 shedding into wastewater. The information gained was used to help monitor, communicate, and manage the COVID-19 pandemic.
Rapid and significant innovations in the field of WBS were realised in parallel in multiple jurisdictions globally. Global collaboration resulted in step-change improvements in multiple aspects of WBS simultaneously, including: health sector leadership; integration with public health surveillance; social and ethical guidance; high-level tactical wastewater catchment sampling strategies; wastewater sampling techniques; transport and storage of samples; laboratory analysis; normalisation and standardisation; data analysis and interpretation; and communication of results.
Innovations in WBS relating to COVID-19 continue to take place, and the benefits of those improvements are now being applied to other targets, such as other pathogens and genes. There are many journal articles and other publications describing these innovations. However, valuable lessons from the COVID-19 pandemic for WBS have not been systematically documented into a practical and readily digestible format.
The purpose of this ‘Cookbook’ is to provide practical and proven ‘recipes’ for WBS for SARS-CoV-2 for two reasons: firstly to help those considering new or extended WBS programmes targeting SARS-CoV-2; and secondly to provide a valuable record of what has been learned from the 2020- 2023 period to help inform WBS programmes to respond to future pandemics and for surveillance of other targets.
The Cookbook is structured into eight parts: 1. Introduction; 2. High-level Overview; 3. Health Sector Perspectives; 4. Methods and Methodologies - From data to action; 5. Methods and Methodologies - Measurement and testing; 6. Methods and Methodologies - Sampling; 6. Methods and Methodologies - Transportation protocols; 8. Conclusions. Multiple authors have contributed to each part, primarily from the European Union, as well as others from the global community of practice.
The Cookbook has been drafted by a global team of authors, with one or more lead authors identified for each chapter and working with their co-authors. The chapters were then subjected to extensive peer review from the external review group. These authors and reviewers are acknowledged in the Cookbook.
Keywords: coronavirus disease, disease surveillance, innovation, public health, report, sampling, standardisation, testing, wastewater
Published in DiRROS: 02.09.2024; Views: 15; Downloads: 9
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52. Guidance on the selection and use of DNA extraction methods : JRC technical reportTheo W. Prins, Wim Broothaerts, Malcolm Burns, Tina Demšar, Sophia Edelmann, Nina Papazova, Verena Peterseil, Isabel Taverniers, 2024, final research report Abstract: DNA extraction is at the forefront of further analytical measurements on DNA targets and affects the downstream results. This report from the European Network of GMO Laboratories (ENGL) provides guidance on the selection and use of fit-for-purpose DNA extraction methods. It focusses on DNA extraction in the context of official controls on the presence and content of genetically modified organisms in food and feed. It provides guidance on protocols and selection support systems, validation approaches, assessment of DNA quality parameters and examples of practical solutions derived from collective experiences. There are many variations on the theme of DNA extraction, but there is no single protocol that works adequately across all food and feed matrices. Before using a new method in the laboratory, or in case of modifications to a protocol, validation or verification is needed to show that a chosen method is fit for purpose for use in routine analysis. This guidance is aimed to help the DNA analysis laboratories in fulfilling the standardisation requirements and support their daily operations.
Keywords: DNA extraction, GMO, detection in food and feed
Published in DiRROS: 02.09.2024; Views: 22; Downloads: 13
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53. Poročilo o preskusu št.: LVG 2024-105 : vzorec št. 2024/00649Barbara Piškur, Patricija Podkrajšek, Špela Hočevar, 2024, expertise, arbitration decision Keywords: varstvo gozdov, morfološke analize, Geosmithia morbida, bolezen tisočerih rakov, qPCR, Juglans, program preiskav
Published in DiRROS: 30.08.2024; Views: 91; Downloads: 0
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54. Metrological evaluation of DNA extraction method effects on the bacterial microbiome and resistome in sputumAleksander Benčič, Nataša Toplak, Simon Koren, Alexandra Bogožalec Košir, Mojca Milavec, Viktorija Tomič, Dane Lužnik, Tanja Dreo, 2024, original scientific article Abstract: Targeted high-throughput sequencing (HTS) has revolutionized the way we look at bacterial communities. It can be used for the species-specific detection of bacteria as well as for the determination of the microbiome and resistome and can be applied to samples from almost any environment. However, the results of targeted HTS can be influenced by many factors, which poses a major challenge for its use in clinical diagnostics. In this study, we investigated the impact of the DNA extraction method on the determination of the bacterial microbiome and resistome by targeted HTS using principles from metrology and diagnostics such as repeatability and analytical sensitivity. Sputum samples spiked with Acinetobacter baumannii, Klebsiella pneumoniae, and Pseudomonas aeruginosa at three different concentrations (103–106 cells/mL) were used. DNA was extracted from each sample on 2 separate days in three replicates each using three different extraction methods based on cetrimonium bromide, magnetic beads, and silica membranes. All three spiked bacteria were detected in sputum, and the DNA extraction method had no significant effect on detection. However, the DNA extraction method had significant effects on the composition of the microbiome and the resistome. The sequencing results were repeatable in the majority of cases. The silica membrane-based DNA extraction kit provided the most repeatable results and the highest diversity of the microbiome and resistome. Targeted HTS has been shown to be a reliable tool for determining the microbiome and resistome; however, the method of DNA extraction should be carefully selected to minimize its impact on the results.
Keywords: targeted high-throughput sequencing, bacterial microbiome, resistome, bacteria detection, DNA extraction, metrology, diagnostics, repeatability
Published in DiRROS: 30.08.2024; Views: 89; Downloads: 46
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55. Degradation processes of two compound layers on nitrided surfaces during the wear test by ‘‘block on hot al cylinder’’Martin Lamut, Jaka Burja, Milan Terčelj, Goran Kugler, David Bombač, 2024, original scientific article Keywords: wear, nitrided steel, hot-work tool steel, thermal fatigue, aluminium extrusion, tool surface degradation
Published in DiRROS: 30.08.2024; Views: 73; Downloads: 86
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58. Poročilo o preskusu št.: LVG 2024-102 : vzorec št. 2024/00610Tine Hauptman, Patricija Podkrajšek, Špela Hočevar, Barbara Piškur, 2024, expertise, arbitration decision Keywords: varstvo gozdov, morfološke analize, Geosmithia morbida, bolezen tisočerih rakov, qPCR, Juglans, program preiskav
Published in DiRROS: 30.08.2024; Views: 87; Downloads: 0
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59. Poročilo o preskusu št.: LVG 2024-101 : vzorec št. 2024/00604Tine Hauptman, Patricija Podkrajšek, Špela Hočevar, Barbara Piškur, 2024, expertise, arbitration decision Keywords: varstvo gozdov, morfološke analize, Geosmithia morbida, bolezen tisočerih rakov, qPCR, Juglans, program preiskav
Published in DiRROS: 30.08.2024; Views: 88; Downloads: 3
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60. Dynamic balance and gait impairments in Parkinson’s disease : novel cholinergic patternsNicolaas I. Bohnen, Uroš Marušič, Stiven Roytman, Rebecca Paalanen, Fotini Michalakis, Taylor Brown, Peter J. H. Scott, Giulia Carli, Roger Albin, Prabesh Kanel, 2024, original scientific article Abstract: The cholinergic system has been implicated in postural deficits, in particular falls, in Parkinson’s disease.
Falls and freezing of gait typically occur during dynamic and challenging balance and gait conditions,
such as when initiating gait, experiencing postural perturbations, or making turns. However, the precise
cholinergic neural substrate underlying dynamic postural and gait changes remains poorly understood.
The aim of this study was to investigate whether brain vesicular acetylcholine transporter binding, as
measured with [18F]-fluoroethoxybenzovesamicolbinding PET, correlates with dynamic gait and balance
impairments in 125 patients with Parkinson’s disease (mean age 66.89±7.71 years) using the abbreviated
Balance Evaluation Systems Test total and its four functional domain sub-scores (anticipatory postural
control, reactive postural control, dynamic gait, and sensory integration). Whole brain false discoverycorrected (P < 0.05) correlations for total abbreviated Balance Evaluation Systems Test scores included
the following bilateral or asymmetric hemispheric regions: gyrus rectus, orbitofrontal cortex, anterior part
of the dorsomedial prefrontal cortex, dorsolateral prefrontal cortex, cingulum, frontotemporal opercula,
insula, fimbria, right temporal pole, mesiotemporal, parietal and visual cortices, caudate nucleus, lateral
and medial geniculate bodies, thalamus, lingual gyrus, cerebellar hemisphere lobule VI, left cerebellar
crus I, superior cerebellar peduncles, flocculus, and nodulus. No significant correlations were found for
the putamen or anteroventral putamen. The four domain-specific sub-scores demonstrated overlapping
cholinergic topography in the metathalamus, fimbria, thalamus proper, and prefrontal cortices but also
showed distinct topographic variations. For example, reactive postural control functions involved the right
flocculus but not the upper brainstem regions. The anterior cingulum associated with reactive postural
control whereas the posterior cingulum correlated with anticipatory control. The spatial extent of
associated cholinergic system changes were least for dynamic gait and sensory orientation functional
domains compared to the anticipatory and reactive postural control functions. We conclude that specific
aspects of dynamic balance and gait deficits in Parkinson’s disease associate with overlapping but also
distinct patterns of cerebral cholinergic system changes in numerous brain regions. Our study also presents
novel evidence of cholinergic topography involved in dynamic balance and gait in Parkinson’s disease
that have not been typically associated with mobility disturbances, such as the right anterior temporal pole,
right anterior part of the dorsomedial prefrontal cortex, gyrus rectus, fimbria, lingual gyrus, flocculus,
nodulus and right cerebellar hemisphere lobules VI and left crus I.
Keywords: Parkinson’s disease, dynamic balance, cholinergic, PET
Published in DiRROS: 30.08.2024; Views: 80; Downloads: 411
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