A Fully Automated Robot for the Preparation of Fungal Samples for FTIR Spectroscopy Using Deep Learning

Manual preparation of fungal samples for Fourier Transform Infrared (FTIR) spectroscopy involves sample washing, homogenization, concentration and spotting, which requires time-consuming and repetitive operations, making it unsuitable for screening studies. This paper presents the design and development of a fully automated robot for the preparation of fungal samples for FTIR spectroscopy. The whole system was constructed based on a previously-developed ultrasonication robot module, by adding a newly-designed centrifuge module and a newly-developed liquid handling module. The liquid handling module consists of a high accuracy electric pipette for spotting and a low accuracy syringe pump for sample washing and concentration. A dual robotic arm system with a gripper connects all of the hardware components. Furthermore, a camera on the liquid handling module uses deep learning to identify the labware settings, which includes the number and positions of well plates and pipette tips. Machine vision on the ultrasonication robot module can detect the sample wells and return the locations to the liquid handling module, which makes the system hand-free for users. Tight integration of all the modules enables the robot to process up to two 96-well microtiter (MTP) plates of samples simultaneously. Performance evaluation shows the deep learning based approach can detect four classes of labware with high average precision, from 0.93 to 1.0. In addition, tests of all procedures show that the robot is able to provide homogeneous sample spots for FTIR spectroscopy with high positional accuracy and spot coverage rate

Oleaginous yeasts respond differently to carbon sources present in lignocellulose hydrolysate

Background Microbial oils, generated from lignocellulosic material, have great potential as renewable and sustainable alternatives to fossil-based fuels and chemicals. By unravelling the diversity of lipid accumulation physiology in different oleaginous yeasts grown on the various carbon sources present in lignocellulose hydrolysate (LH), new targets for optimisation of lipid accumulation can be identified. Monitoring lipid formation over time is essential for understanding lipid accumulation physiology. This study investigated lipid accumulation in a variety of oleaginous ascomycetous and basidiomycetous strains grown in glucose and xylose and followed lipid formation kinetics of selected strains in wheat straw hydrolysate (WSH). Results Twenty-nine oleaginous yeast strains were tested for their ability to utilise glucose and xylose, the main sugars present in WSH. Evaluation of sugar consumption and lipid accumulation revealed marked differences in xylose utilisation capacity between the yeast strains, even between those belonging to the same species. Five different promising strains, belonging to the species Lipomyces starkeyi, Rhodotorula glutinis, Rhodotorula babjevae and Rhodotorula toruloides, were grown on undiluted wheat straw hydrolysate and lipid accumulation was followed over time, using Fourier transform-infrared (FTIR) spectroscopy. All five strains were able to grow on undiluted WSH and to accumulate lipids, but to different extents and with different productivities. R. babjevae DVBPG 8058 was the best-performing strain, accumulating 64.8% of cell dry weight (CDW) as lipids. It reached a culture density of 28 g/L CDW in batch cultivation, resulting in a lipid content of 18.1 g/L and yield of 0.24 g lipids per g carbon source. This strain formed lipids from the major carbon sources in hydrolysate, glucose, acetate and xylose. R. glutinis CBS 2367 also consumed these carbon sources, but when assimilating xylose it consumed intracellular lipids simultaneously. Rhodotorula strains contained a higher proportion of polyunsaturated fatty acids than the two tested Lipomyces starkeyi strains. Conclusions There is considerable metabolic diversity among oleaginous yeasts, even between closely related species and strains, especially when converting xylose to biomass and lipids. Monitoring the kinetics of lipid accumulation and identifying the molecular basis of this diversity are keys to selecting suitable strains for high lipid production from lignocellulose

Montana Kaimin, January 30, 2008

Student newspaper of the University of Montana, Missoula.https://scholarworks.umt.edu/studentnewspaper/6138/thumbnail.jp

Assessment of genetically modified maize Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 for food and feed uses, import and processing under Regulation (EC) No 1829/2003 (application EFSA-GMO-DE-2018-149)

Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 was produced by conventional breeding of the GM maize events Bt11, MIR162, MIR604, MON 89034, 5307 and GA21. Accordingly, Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 maize produces the transgenic proteins in the individual GM maize events (Cry1Ab, PAT, Vip3Aa20, PMI, mCry3A, MIR604 PMI, Cry1A.105, Cry2Ab2, eCry3.1Ab and mEPSPS). Event Bt11 maize expresses the insecticidal protein Cry1Ab that protects against feeding damage caused by certain lepidopteran pests and the phosphinothricin acetyltransferase (PAT) protein for weed control by providing tolerance to herbicide products containing glufosinate ammonium. Event MIR162 maize expresses the insecticidal protein Vip3Aa20 that protects against feeding damage caused by certain lepidopteran pests and the PMI protein which enables transformed plant cells to utilise mannose as a primary carbon source and therefore used as a selectable marker in the development of the MIR162 maize. Event MIR604 maize expresses the insecticidal protein mCry3A that protects against feeding damage caused by certain coleopteran pests and the MIR604 PMI protein which enables transformed plant cells to utilise mannose as a primary carbon source and therefore used as a selectable marker in the development of the MIR604 maize. Event MON 89034 maize expresses the insecticidal proteins Cry1A.105 and Cry2Ab2 that protect against feeding damage caused by certain lepidopteran pests. Event 5307 maize expresses the insecticidal protein eCry3.1Ab that protects against feeding damage caused by certain coleopteran pests and the PMI protein which enables transformed plant cells to utilise mannose as a primary carbon source and therefore used as a selectable marker in the development of the 5307 maize. Event GA21 expresses the double-mutated 5-enolpyruvylshikimate-3-phosphate synthase enzyme (mEPSPS) for weed control by providing tolerance to herbicide products containing glyphosate.The scientific documentation provided in the application for genetically modified maize Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 is adequate for risk assessment, and in accordance with EFSA guidance on risk assessment of genetically modified plants for use in food or feed. The VKM GMO panel does not consider the introduced modifications in maize Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 to imply potential specific health or environmental risks in Norway, compared to EU-countries The EFSA opinion is adequate also for Norwegian considerations. Therefore, a full risk assessment of maize Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 was not performed by the VKM GMO Panel.Assessment of genetically modified maize Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 for food and feed uses, import and processing under Regulation (EC) No 1829/2003 (application EFSA-GMO-DE-2018-149)publishedVersio

Rhodotorula kratochvilovae CCY 20-2-26-The Source of Multifunctional Metabolites

Multifunctional biomass is able to provide more than one valuable product, and thus, it is attractive in the field of microbial biotechnology due to its economic feasibility. Carotenogenic yeasts are effective microbial factories for the biosynthesis of a broad spectrum of biomolecules that can be used in the food and feed industry and the pharmaceutical industry, as well as a source of biofuels. In the study, we examined the effect of different nitrogen sources, carbon sources and CN ratios on the co-production of intracellular lipids, carotenoids, beta-glucans and extracellular glycolipids. Yeast strain R. kratochvilovae CCY 20-2-26 was identified as the best co-producer of lipids (66.7 +/- 1.5% of DCW), exoglycolipids (2.42 +/- 0.08 g/L), beta-glucan (11.33 +/- 1.34% of DCW) and carotenoids (1.35 +/- 0.11 mg/g), with a biomass content of 15.2 +/- 0.8 g/L, by using the synthetic medium with potassium nitrate and mannose as a carbon source. It was shown that an increased C/N ratio positively affected the biomass yield and production of lipids and beta-glucans

Assessment of genetically modified maize Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 for food and feed uses, import and processing under Regulation (EC) No 1829/2003 (application EFSA-GMO-DE-2018-149)

Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 was produced by conventional breeding of the GM maize events Bt11, MIR162, MIR604, MON 89034, 5307 and GA21. Accordingly, Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 maize produces the transgenic proteins in the individual GM maize events (Cry1Ab, PAT, Vip3Aa20, PMI, mCry3A, MIR604 PMI, Cry1A.105, Cry2Ab2, eCry3.1Ab and mEPSPS). Event Bt11 maize expresses the insecticidal protein Cry1Ab that protects against feeding damage caused by certain lepidopteran pests and the phosphinothricin acetyltransferase (PAT) protein for weed control by providing tolerance to herbicide products containing glufosinate ammonium. Event MIR162 maize expresses the insecticidal protein Vip3Aa20 that protects against feeding damage caused by certain lepidopteran pests and the PMI protein which enables transformed plant cells to utilise mannose as a primary carbon source and therefore used as a selectable marker in the development of the MIR162 maize. Event MIR604 maize expresses the insecticidal protein mCry3A that protects against feeding damage caused by certain coleopteran pests and the MIR604 PMI protein which enables transformed plant cells to utilise mannose as a primary carbon source and therefore used as a selectable marker in the development of the MIR604 maize. Event MON 89034 maize expresses the insecticidal proteins Cry1A.105 and Cry2Ab2 that protect against feeding damage caused by certain lepidopteran pests. Event 5307 maize expresses the insecticidal protein eCry3.1Ab that protects against feeding damage caused by certain coleopteran pests and the PMI protein which enables transformed plant cells to utilise mannose as a primary carbon source and therefore used as a selectable marker in the development of the 5307 maize. Event GA21 expresses the double-mutated 5-enolpyruvylshikimate-3-phosphate synthase enzyme (mEPSPS) for weed control by providing tolerance to herbicide products containing glyphosate.The scientific documentation provided in the application for genetically modified maize Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 is adequate for risk assessment, and in accordance with EFSA guidance on risk assessment of genetically modified plants for use in food or feed. The VKM GMO panel does not consider the introduced modifications in maize Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 to imply potential specific health or environmental risks in Norway, compared to EU-countries The EFSA opinion is adequate also for Norwegian considerations. Therefore, a full risk assessment of maize Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 was not performed by the VKM GMO Panel.Assessment of genetically modified maize Bt11 x MIR162 x MIR604 x MON 89034 x 5307 x GA21 for food and feed uses, import and processing under Regulation (EC) No 1829/2003 (application EFSA-GMO-DE-2018-149)publishedVersio

Assessment of genetically modified maize DP915635 for food and feed uses, import and processing under Regulation (EC) No 1829/2003 (application EFSA-GMO-NL-2020-172)

The Norwegian Scientific Committee for Food and Environment (VKM) has assessed an application for approval of the genetically modified maize DP915635 for food and feed uses, import and processing in the EU. In accordance with an assignment specified by the Norwegian Food Safety Authority (NFSA) and the Norwegian Environment Agency (NEA), VKM assesses whether genetically modified organisms (GMOs) intended for the European market can pose risks to human or animal health, or the environment in Norway. VKM assesses the scientific documentation regarding GMO applications seeking approval for use of GMOs as food and feed, processing, or cultivation. The EU Regulation 1829/2003/EC (Regulation) covers living GMOs that fall under the Norwegian Gene Technology Act, as well as processed food and feed from GMOs (dead material) that fall under the Norwegian Food Act. The regulation is currently not part of the EEA agreement or implemented in Norwegian law. Norway conducts its own assessments of GMO applications in preparation for the possible implementation of the Regulation. In accordance with the assignment by NFSA and NEA, VKM assesses GMO applications during scientific hearings initiated by the European Food Safety Authority (EFSA), as well as after EFSA has published its own risk assessment of a GMO, up until EU member countries vote for or against approval in the EU Commission. The assignment is divided into three stages. (link) Maize DP915635 DP915635 is a genetically modified maize that expresses the insecticidal protein IPD079Ea for control of corn rootworm pests, the enzyme phosphinothricin acetyltransferase (PAT) for tolerance to glufosinate-ammonium herbicides, and the enzyme phosphomannose isomerase (PMI) that was used as a selectable marker during development. The scientific documentation provided in the application for DP915635 maize is adequate for risk assessment, and in accordance with EFSA guidance on risk assessment of genetically modified plants for use in food or feed. The VKM GMO panel does not consider the introduced modifications in DP915635 maize to imply potential specific health or environmental risks in Norway, compared to EU-countries. The EFSA scientific Opinion is adequate also for Norwegian conditions. Therefore, a full risk assessment of DP915635 maize was not performed by the VKM GMO Panel. About the assignment: In stage 1, VKM shall assess the health and environmental risks of the genetically modified organism and derived products in connection with the EFSA scientific hearing of GMO applications. VKM shall review the scientific documentation that the applicant has submitted and possibly provide comments to EFSA. VKM must also consider: i) whether there are specific Norwegian conditions that could give other risks in Norway than those mentioned in the application, ii) whether the Norwegian diet presents a different health risk for the Norwegian population should the GMO be approved, compared to the European population, and iii) risks associated with co-existence with conventional and/or ecologic production of plants for GMOs seeking approval for cultivation. Relevant measures to ensure co-existence must also be considered. In stage 2, VKM shall assess whether comments from Norway have been satisfactorily answered by EFSA. In addition, VKM shall assess whether comments from other countries imply need for further follow-up. If EFSAs response to Norwegian comments is not satisfactory, or comments by other countries imply the need for further follow-up, VKM shall in stage 3 perform a risk assessment of these conditions, including conditions specific to Norway.Assessment of genetically modified maize DP915635 for food and feed uses, import and processing under Regulation (EC) No 1829/2003 (application EFSA-GMO-NL-2020-172)publishedVersio

Assessment of genetically modified soybean MON 87701 × MON 89788 for food and feed uses, import and processing under Regulation (EC) No 1829/2003 (renewal application EFSAGMO-RX-022)

publishedVersio

Assessment of genetically modified cotton COT102 for food and feed uses, import and processing under Regulation (EC) No 1829/2003 (application EFSA-GMO-DE-2017-141)

publishedVersio

Assessment of genetically modified maize 94804 for food and feed uses, import and processing under Regulation (EC) No 1829/2003 (application GMFF-2022-10651/EFSA-Q-2023- 00106 (182))

The Norwegian Scientific Committee for Food and Environment (VKM) has assessed an application for approval of the genetically modified maize MON 94804 for food and feed uses, import and processing in the EU. In accordance with an assignment specified by the Norwegian Food Safety Authority (NFSA) and the Norwegian Environment Agency (NEA), VKM assesses whether genetically modified organisms (GMOs) intended for the European market can pose risks to human or animal health, or the environment in Norway. VKM assesses the scientific documentation regarding GMO applications seeking approval for use of GMOs as food and feed, processing, or cultivation. The EU Regulation 1829/2003/EC (Regulation) covers living GMOs that fall under the Norwegian Gene Technology Act, as well as processed food and feed from GMOs (dead material) that fall under the Norwegian Food Act. The regulation is currently not part of the EEA agreement or implemented in Norwegian law. Norway conducts its own assessments of GMO applications in preparation for the possible implementation of the Regulation. In accordance with the assignment by NFSA and NEA, VKM assesses GMO applications during scientific hearings initiated by the European Food Safety Authority (EFSA), as well as after EFSA has published its own risk assessment of a GMO, up until EU member countries vote for or against approval in the EU Commission. The assignment is divided into three stages. (link) Maize MON 94804 MON 94804 is a genetically modified maize developed via Agrobacterium tumefaciens -mediated transformation of maize cells. Maize MON 94804 expresses a micro-RNA (miRNA) transcript, that leads to reduced levels of the hormone gibberellin in the plant. Gibberellin is involved in, i.a., plant stem elongation. Thus, maize MON 94804 plants grow shorter than other maize. The scientific documentation provided in the application for maize MON 94804 is adequate for risk assessment, and in accordance with EFSA guidance on risk assessment of genetically modified plants for use in food or feed. The VKM GMO panel does not consider the introduced modifications in maize MON 94804 to imply potential specific health or environmental risks in Norway, compared to EU-countries. The EFSA scientific Opinion is adequate also for Norwegian conditions. Therefore, a full risk assessment of maize MON 94804 was not performed by the VKM GMO Panel.Assessment of genetically modified maize 94804 for food and feed uses, import and processing under Regulation (EC) No 1829/2003 (application GMFF-2022-10651/EFSA-Q-2023- 00106 (182))publishedVersio