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Nutrition is one of the main topics of the society in general. In modern times, it became even more important due to increased rate of metabolic disorders, such as obesity, diabetes type 2, and cardiovascular diseases. There are various possible causes to these disorders, however the great proportion is a consequence of bad dietary habits (e.g., overeating, poor quality diet) and lack of physical activity. To stop this trend, some prevention measures can be taken such as wide screening of the population and personalized dietary approach. In this work, we performed large-scale validation of the portable and affordable mass spectrometer for food impact assessment from volatile organic compounds (VOCs) in breath. Validation was done as a pilot study among 6 different population groups: people with overweightness, people with obesity, people with diabetes type II, people with cardiovascular disease, people with poor quality diet and professional athletes. More than 150 participants were recruited from 5 European countries – UK, Greece, Germany, Belgium, and Portugal with informed consents and ethical approvals. Food impact was assessed by monitoring the concentrations of several VOCs (acetone, ethanol, isoprene, and n-pentane) related to macronutrients metabolism in exhaled breath samples. The analysis was done using the newly developed membrane inlet mass spectrometer (MIMS). Concentration levels for selected breath VOCs were determined in samples collected before the meal and after the meal. The experimental data was compared against the data acquired via the questionnaire to discover possible relationships between VOCs concentration levels and meal consumption trends among specific population groups. The results will be incorporated into the mobile application for personalized nutrition and analyzed by the experts.

Food metabolism is very complex, and impact of specific diet on human wellness can be difficult to reveal. Due to its non-invasive nature, breath analysis is getting more engaged in nutritional research. However, extensive research among different population groups is still required. To provide accurate diagnostic and diet recommendations, an individual approach is required. For wide screening accomplishment, it is necessary to establish novel, portable and affordable solutions to complement conventional diagnostic techniques. Therefore, a novel sensor for breath VOCs analysis was utilized in this research on different population groups. Exhaled breath samples were collected from people with diabetes in Germany and people with cardiovascular diseases in Belgium together with informed consents and ethical approvals. Along with the samples, all the participants provided information about their lifestyle via short questionnaire. Samples were collected in 1L Tedlar bags with one sample being taken before the meal and one 2h after the meal. Key compounds that were monitored were breath acetone, ethanol, isoprene, and n-pentane. They were analysed using a portable membrane inlet mass spectrometer (MIMS) according to a previously established method [1]. Quantified VOCs levels in people with diabetes and people with cardiovascular diseases were compared with those from healthy participants, previously obtained using the same instrument and method [1]. ANOVA statistical test was used for determination of possible statistically significant differences among examined groups of participants and VOCs levels changes upon the food consumption. Involvement of more participants from different populations will enable database set for future VOC sensor predictions. [1] Jakšić, M., Mihajlović, A., Vujić, D., Giannoukos, S., Brkić, B.Membrane inlet mass spectrometry method for food intake impact assessment on specific volatile organic compounds in exhaled breath.Anal Bioanal Chem(2022). https://doi.org/10.1007/s00216-022-04168-3

Introduction: It is well known that exhaled breath composition is instant reflection of the organism since volatile metabolites tend to move from the blood into the air in the lungs. Therefore, breatholomics is becoming very popular as а new non-invasive screening diagnostic tool. This research brings a portable, low-cost screening sensor prototype based on membrane-inlet mass spectrometry (MIMS) for food impact assessment, by tracking changes of volatile organic compounds (VOCs) in exhaled breath. Analytes of interest are breath VOCs that are related to the metabolism of the main food constituents. Methods: 150 exhaled breath samples were collected from 50 adult healthy volunteers with informed consents and ethical approval completed (Approval No. 2021-01-3/70-1). Each participant provided a sample before meal (after 12h fasting period), 60 min and 120 min after the meal. All samples were analyzed using MIMS system for selected VOC mass fragments – m/z 58 for acetone, m/z 42 for n-pentane, m/z 45 for ethanol and m/z 67 for isoprene. Sample introduction was done using PDMS sheet membrane probe heated at 70◦C. VOC gas standards were prepared by static dilution technique and used for quantification. Results: Significant change (more than 10% increment or decrement) upon meal consumption was observed in ~85% of participants for acetone and ethanol, ~70% for n-pentane and ~50% for isoprene. Changes in selected analytes’ signals were compared for 60min and 120min after the meal, and greater change was observed 120 min after the meal for all analytes, except for ethanol. Obtained mean values for acetone, ethanol, n-pentane, and isoprene respectively were: 809 ppb, 488 ppb, 30 ppb and 70 ppb in samples before meal, 875 ppb, 532 ppb, 29 ppb and 68 ppb in samples collected 60 min after the meal and 559 ppb, 474 ppb, 27 ppb and 66 ppb in samples collected 120 min after the meal. According to the results obtained, it can be said that MIMS system used in this study could be employed in food impact assessment by analyzing breath VOCs. It has been established that major food impact can be monitored 120 min after the meal for analyzed mass fragments. Additionally, obtained mean values were ranked in expected order of magnitude and not surprisingly – the most significant food impact was observed for acetone. Further research will include more participants with different nutritional and health backgrounds, which will expand the knowledge in this area. Novel Aspect: Employment of portable MIMS system in food impact assessment offers a great advantage for possible preventive screening analyses that would provide an alert for nutritional imbalances.

Background: Champignon mushroom production is strongly influenced by the occurrence of pathogenic bacteria and fungi causing different kinds of diseases which can significantly lower the yield. Green mold disease is one of the most aggressive diseases of champignon mushrooms. In champignon production, green mold disease is connected to fungal species belonging to genus Trichoderma. The most aggressive are T. aggressivum f. europaeum (mainly present in Europe) and T. aggressivum f. aggressivum (mainly present in North America), but also T. harzianum. Objectives: In this research we aimed to explore the efficiency of DNA metabarcoding approach to detect Trichoderma species present in compost and casing soil during champignon cultivation. Methods: Genomic DNA extracted from compost and casing soil was used as a template for amplification and sequencing of an ITS2 fragment. High-throughput sequencing was performed on the Illumina NovaSeq 6000 platform. Raw sequence data was processed using next-generation microbiome bioinformatics platform QIIME2. In total, 332 operational taxonomic units were defined and the trend of succession in fungal communities of compost and casing soil were detected between different time dependent sample series. Results: We identified eight Trichoderma species among which T. harzianum and T. aggressivum. The first was detected in all sample series, while the second was detected only in casing soil samples. Using DNA metabarcoding it was possible to detect aggressive Trichoderma species even before the appearance of visible green mold. However, there was no clear pattern in Trichoderma spp. detection among time dependent sample series.

Introduction Nutritional balance is among the major concerns of modern people, as it is known that diet directly impacts the overall state of the human body. To choose the optimal diet for everyone, a personalized approach is needed. The purpose of this study was to validate a sensor for food impact assessment by analyzing the levels of volatile organic compounds (VOCs) in human breath. It aims to establish a prototype for future auxiliary technique for nutritional status clarification. Breath research has been very popular in the last 10 years, mainly due to its non-invasive diagnostic potential. As the proposed sensor does not require laboratory conditions, its application could be widespread due to fast analysis, portability, and affordability. Methods This pilot study recruited various groups of participants from several European countries, with informed consents and ethical approvals. The groups included overweight participants, those with obesity, and those who have poor dietary habits. Food impact assessment was conducted by monitoring levels of several VOCs related to macronutrients metabolism in exhaled breath samples using the new portable sensor based on membrane inlet mass spectrometry (MIMS). Each participant provided a sample before standardized meal (after 12h food restraint) and 120 min after the meal. Also, information about participants’ lifestyle was collected via a short questionnaire. Samples were collected in 1L single-use Tedlar®bags and analyzed using the MIMS system with PDMS sheet membrane probe. Preliminary Data Functionality of the new portable breath VOC sensor was verified by examining several analytical parameters in our preliminary research study on a group of healthy participants (not published yet). That pre-pilot study showed that the examined MIMS system could be used successfully for breath VOCs analysis, and for monitoring the changes in VOCs levels upon food consumption. A large-scale pilot study employing the new method on more population groups was the natural continuation of the pre-pilot study. Concentration levels for selected breath VOCs (acetone, ethanol, isoprene and n-pentane) were determined in samples before the meal (BM) and after the meal (AM). Additionally, qualitative and quantitative aspects of food impact assessment were determined and evaluated. The qualitative assessment provided information about whether a specific breath VOC increased or decreased as a consequence of food ingestion. Furthermore, quantification of the food impact was established via comparison factor. This factor represents the ratio of VOC levels after and before the meal, i.e., AM/BM for the same participant. Comparison factor served to express the intensity of the change in VOCs signal. More than 10% of signal increment or decrement was considered as food impact. Additionally, the experimentally obtained results were examined against the data collected via the questionnaire in order to discover possible correlations between VOCs changes and specific population group or their lifestyle habits. The results obtained in this pilot study support the fact that proposed VOC sensor can be utilized in breath research and contribute to the enrichment of the breath research area. Novel Aspect Portable MIMS sensor for detection of VOCs showed potential to become fast screening methodology for nutritional status diagnostics.

from the BelBi2021 conference