Developmental Validation of Short Tandem Repeat Reagent Kit for Forensic DNA Profiling of Canine Biological Material

Aim To develop a reagent kit that enables multiplex polymerase chain reaction (PCR) amplification of 18 short tandem repeats (STR) and the canine sex-determining Zinc Finger marker. Methods Validation studies to determine the robustness and reliability in forensic DNA typing of this multiplex assay included sensitivity testing, reproducibility studies, intra- and inter-locus color balance studies, annealing temperature and cycle number studies, peak height ratio determination, characterization of artifacts such as stutter percentages and dye blobs, mixture analyses, species- specificity, case type samples analyses and population studies. Results The kit robustly amplified domesticated dog samples and consistently generated full 19-locus profiles from as little as 125 pg of dog DNA. In addition, wolf DNA samples could be analyzed with the kit. Conclusion The kit, which produces robust, reliable, and reproducible results, will be made available for the forensic research community after modifications based on this study’s evaluation to comply with the quality standards expected for forensic casework

Modernisation of agricultural statistics : Final report

The “Modernisation of agricultural statistics” project had three main goals: (1) modernisation of the statistical register of agricultural and horticultural enterprises (Mapu), (2) identification of new data sources that use agricultural statistics, and (3) the better utilisation of farm related geospatial data to define the location of farms. In addition, an international seminar was held in Poland during the project in cooperation with other countries carrying out a similar project and with representatives of Eurostat. The project was divided into work packages in accordance with the main goals. The first work package included the modernisation of Mapu. This work package was further divided into two main tasks based on data sources: administrative data and direct data collection. Before the concrete modernisation of systems and databases, we identified how well the definition of a farm used in Mapu corresponds with the definition of a farm used in EU statistics (Article 2 of regulation (EU) 2018/1019 on integrated farm statistics). In practice, the definition of a farm entity used in Mapu in accordance with the Integrated Administration and Control System (IACS) corresponds well with the definition used in EU statistics. The most significant difference is that Mapu mainly does not include fur farming, reindeer husbandry and beekeeping. Apart from fur farming, the significance of these sectors is low, and this project does not recommend that enterprises engaged in these sectors be added to Mapu. Fur farms are included in the Business Register of Statistics Finland, which can be used to produce statistics of fur farms. If statistical obligations related to fur farming expand later, for example, as a result of the SAIO regulation, adding fur farms to Mapu should be reassessed. Mapu’s databases and related data flows were modernised during the project. The removal of direct database rights to datasets of agricultural administration for reasons related to information security was a significant change regarding administrative data. The Hansolo loading platform was built as a new solution. The administrative data required for agricultural statistics is copied regularly in Hansolo, from which data is available for Mapu and agricultural statistics. In addition to databases and data flows, the data content was modernised. Adding the contact details and business ID of farm parties, such as farmers, to the data content was the most significant addition. The data system of the administrative body of rural industries, belonging to IACS, is the primary data source. Data about animals is obtained from animal registers. In other words, units (farms and horticultural enterprises) in Mapu are the same as units in IACS. Some data in Mapu can be obtained through direct data collection processes in conjunction with the survey for horticultural statistics. During this project, the data collection application, databases and data flows of this survey were modernised regarding all process phases, ranging from data collection to publication. As a new feature in data collection for horticultural statistics, farmers were able to log in to the online application through strong electronic identification using their banking codes, for example. As a result, it was not necessary to send usernames and passwords separately to respondents. In 2020, the data collection application was developed further by introducing an authorisation feature. In practice, this means that farmers can authorise another person to respond to the survey on their behalf. In the second work package, we identified the broader use of data about farm coordinates to define the location of farms in agricultural statistics. The aim is to present data on maps better than before, while considering the data privacy of individual farms. In the third work package, we examined what new data sources could be used more effectively than at present in agricultural statistics. According to the examination, data in the parcel databank about the use of crop protection products apparently represented the actual use of products better than data collected through the current data collection process. The use of parcel specific data obtained from farms for various purposes is developing and, as a result, parcel data may also become available as source material for statistics by new electronic means. In addition, we developed a method to calculate the field-specific monoculture area. Compared with the previous method version, the use of the QuantumGIS program was a new factor. It helped to accurately define the area used for a single crop in consecutive years, also when the crop was only grown in part of the parcel in question during the three years under examination. The method for calculating the monoculture area developed in this project will be used in the 2023 IFS data collection process to produce data about crop rotation. Considering the future of agricultural statistics, it will be interesting to see how new production sectors, such as insect farming, will possibly be included in the statistics. This also applies to questions related to production methods: in the future, food may be produced by enterprises other than conventional farms, such as by enterprises engaged in vertical farming. Environmental issues have steered agricultural activities in the past and will continue to do so during the next few years. For example, sequestering carbon in farmland may expand beyond perennial grasslands and fallows. Data production related to products sold, combined with statistics of environmental impact arising from agriculture, presents a challenge in agricultural statistics. Growing and new statistical needs, together with fewer resources, will steer statistical production during the next few years towards the use of new data sources and the identification of data production that is “sufficiently good” in terms of quality and finances. Modernisation of agricultural statistics co-funded by the Eurostat (Finland Grant agreement 2018.0212). This publication is the final report of the project (20.8.2020)

Scenario-based assessment of fecal pathogen sources affecting bathing water quality: novel treatment options to reduce norovirus and Campylobacter infection risks

Wastewater discharge and runoff waters are significant sources of human and animal fecal microbes in surface waters. Human-derived fecal contamination of water is generally estimated to pose a greater risk to human health than animal fecal contamination, but animals may serve as reservoirs of zoonotic pathogens. In this study, quantitative microbial risk assessment (QMRA) tools were used to evaluate the hygienic impact of sewage effluents and runoff water from municipalities and animal farms on surface and bathing waters. The human-specific microbial source tracking (MST) marker HF183 was used to evaluate the dilution of fecal pathogens originating from the sewage effluent discharge to the downstream watershed. As novel risk management options, the efficiency of UV-LED disinfection and wetland treatment as well as biochar filtration was tested on-site for the contamination sources. According to the dilution pattern of the MST marker HF183, microbes from wastewater were diluted (2.3–3.7 log10) in the receiving waters. The scenario-based QMRA revealed, that the health risks posed by exposure to human-specific norovirus GII and zoonotic Campylobacter jejuni during the bathing events were evaluated. The risk for gastroenteritis was found to be elevated during wastewater contamination events, where especially norovirus GII infection risk increased (1–15 cases per day among 50 bathers) compared with the business as usual (BAU) situation (1 case per day). The noted C. jejuni infection risk was associated with animal farm contamination (1 case per day, versus 0.2–0.6 cases during BAU). Tertiary treatment of wastewater with wetland treatment and UV-LED disinfection effectively reduced the waterborne gastroenteritis risks associated with bathing. Based on the experiences from this study, a QMRA-based approach for health risk evaluations at bathing sites can be useful and is recommended for bathing site risk assessments in the future. In case of low pathogen numbers at the exposure sites, the MST marker HF183 could be used as a pathogen dilution coefficient for the watershed under evaluation. The full-scale implementation of novel tertiary treatment options at wastewater treatment plants (WWTPs) as well as on-site runoff water treatment options should be considered for infection risk management at locations where scenario-based QMRA implies elevated infection risks

A Synthesis of Marine Monitoring Methods With the Potential to Enhance the Status Assessment of the Baltic Sea

Highlights - We rated novel methods regarding their ability to improve the Baltic Sea monitoring. - Methods were assessed with respect to their costs and applicability. - All methods can potentially increase data resolution or monitor novel ecosystem elements. - We recommend several novel methods for the Baltic status assessment.A multitude of anthropogenic pressures deteriorate the Baltic Sea, resulting in the need to protect and restore its marine ecosystem. For an efficient conservation, comprehensive monitoring and assessment of all ecosystem elements is of fundamental importance. The Baltic Marine Environment Protection Commission HELCOM coordinates conservation measures regulated by several European directives. However, this holistic assessment is hindered by gaps within the current monitoring schemes. Here, twenty-two novel methods with the potential to fill some of these gaps and improve the monitoring of the Baltic marine environment are examined. We asked key stakeholders to point out methods likely to improve current Baltic Sea monitoring. We then described these methods in a comparable way and evaluated them based on their costs and applicability potential (i.e., possibility to make them operational). Twelve methods require low to very low costs, while five require moderate and two high costs. Seventeen methods were rated with a high to very high applicability, whereas four methods had moderate and one low applicability for Baltic Sea monitoring. Methods with both low costs and a high applicability include the Manta Trawl, Rocket Sediment Corer, Argo Float, Artificial Substrates, Citizen Observation, Earth Observation, the HydroFIA®pH system, DNA Metabarcoding and Stable Isotope Analysis

α5β1 integrin recycling promotes Arp2/3-independent cancer cell invasion via the formin FHOD3

Invasive migration in 3D extracellular matrix (ECM) is crucial to cancer metastasis, yet little is known of the molecular mechanisms that drive reorganization of the cytoskeleton as cancer cells disseminate in vivo. 2D Rac-driven lamellipodial migration is well understood, but how these features apply to 3D migration is not clear. We find that lamellipodia-like protrusions and retrograde actin flow are indeed observed in cells moving in 3D ECM. However, Rab-coupling protein (RCP)-driven endocytic recycling of α5β1 integrin enhances invasive migration of cancer cells into fibronectin-rich 3D ECM, driven by RhoA and filopodial spike-based protrusions, not lamellipodia. Furthermore, we show that actin spike protrusions are Arp2/3-independent. Dynamic actin spike assembly in cells invading in vitro and in vivo is regulated by Formin homology-2 domain containing 3 (FHOD3), which is activated by RhoA/ROCK, establishing a novel mechanism through which the RCP–α5β1 pathway reprograms the actin cytoskeleton to promote invasive migration and local invasion in vivo

Ratkaisuja Suomen ruokastrategiaan. FOOD-ohjelman tietopaketti päättäjille

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