The information on microbiota composition in the human gastrointestinal tract predominantly originates from the analyses of human faeces by application of next generation sequencing (NGS). However, the detected composition of the faecal bacterial community can be affected by various factors including experimental design and procedures. This study evaluated the performance of different protocols for collection and storage of faecal samples (native and OMNIgene.GUT system) and bacterial DNA extraction (MP Biomedicals, QIAGEN and MO BIO kits), using two NGS platforms for 16S rRNA gene sequencing (Ilumina MiSeq and Ion Torrent PGM). OMNIgene.GUT proved as a reliable and convenient system for collection and storage of faecal samples although favouring Sutterella genus. MP provided superior DNA yield and quality, MO BIO depleted Gram positive organisms while using QIAGEN with OMNIgene.GUT resulted in greatest variability compared to other two kits. MiSeq and IT platforms in their supplier recommended setups provided comparable reproducibility of donor faecal microbiota. The differences included higher diversity observed with MiSeq and increased capacity of MiSeq to detect Akkermansia muciniphila, [Odoribacteraceae], Erysipelotrichaceae and Ruminococcaceae (primarily Faecalibacterium prausnitzii). The results of our study could assist the investigators using NGS technologies to make informed decisions on appropriate tools for their experimental pipelines.

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Introduction: The primary aims of this study were to evaluate the changes in dietary behavior among the Croatian adult population during the COVID-19 outbreak and to explore the impact of confinement on cooking habits.Methods: The study was based on results from COVIDiet_Int cross-sectional study—a part of COVIDiet project (NCT04449731). A self-administered online questionnaire was used to assess the frequency of food consumption, eating habits, and sociodemographic information. A total number of 4,281 participants (80.5% females and 19.4% males) completed the questionnaire.Results: The Mediterranean Diet Adherence Screener (MEDAS) score before the confinement was 5.02 ± 1.97, while during the confinement, the MEDAS score increased to 5.85 ± 2.04. Participants who had higher adherence to the Mediterranean diet (MedDiet) during the confinement were mostly females (88.8%), aged between 20 and 50 years, with the highest level of education (66.3%) and normal BMI (70.6%). The majority of participants maintained their dietary behavior as it was before COVID-19 confinement, while 36.9% decreased their physical activity. Participants with higher MEDAS score were more eager to increase their physical activity. Additionally, higher median values of MEDAS score were noted for participants with body mass index values below 24.9 kg/m2 (6.0 vs. 5.0 for participants with BMI above 25 kg/m2). Participants in all residence places increased their cooking frequency during the confinement (53.8%), which was associated with an increase in vegetables, legumes, as well as fish and seafood consumption.Conclusions: According to our findings, Croatian adults exhibited medium adherence to the MedDiet during the COVID-19 confinement. The results suggest that cooking frequency could be positively associated with overall dietary quality, which is of utmost importance in these demanding times.

Challenging periods like the COVID-19 pandemic require fast and efficient adaptations of the healthcare system. It is vital that every patient has access to nutritional care as a part of primary healthcare services, even if social distancing measures are adopted. Therefore, we propose a simple remote nutritional screening tool and practical guidance for nutritional care in primary practice, and their implementation into telemedicine processes and digital platforms suitable for healthcare providers. The acronym for the tool is R-MAPP, as for Remote e Malnutrition APP, while the tool will be available also as an app. This protocol consists of two simple validated clinical tools for identifying nutritional risk and loss of muscle mass and function eMalnutrition Universal Screening Tool ('MUST') and SARC-F (5-item questionnaire: Strength, Assistance with walking, Rise from a chair, Climb stairs and Falls) -and additional practical guidance on nutritional interventions for family physicians.