National Research Institute for Agriculture, Food and Environment

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    20th International Symposium on Field- and Flow-Based Separations

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    Milk contains two types of proteins: Caseins and whey proteins. There are four caseins: α/β/γ/κ caseins that formed micelles in milk. Whey proteins are globular proteins between 12 and 200 kDa: α-lactalbumin; β-lactoglobulin; Serum albumin; Immunoglobulin and Lactoferrin. Industrials fractionate milk to produce whey protein and casein isolate to make ingredients for the formulation of dairy products. Whey protein isolate can be heated to functionalize or pasteurize them. The objective of these three studies are to characterize the isolates of whey proteins and the impact of heating treatment on whey protein fractions and formulations using different FFF techniques: Asymmetrical-FFF; 2D AsFlFFF-RPLC and Centrifugal-FFF Whey protein isolate heated at high temperature were studied by Centrifugal-FFF coupled with MALS and RI. Aggregates with the lowest density and size elute in the void peak and represent 51% of the mass fraction. For the bigger one, there are two populations: one smaller than 1 micrometer and one higher, up to 4.5 micrometers. For size determination, the biggest population fractionated has been collected and analyzed by DLS in batch. Infant milk formulation without fat was investigated by Asymmetrical Flow FFF coupled with MALS; QELS and RI. The formulation contains bovine milk and isolate whey protein fractions. The technique separates the non-aggregated whey protein to the casein micelles. The addition of lactoferrin partially disintegrated the casein micelles. The formulations are heated at 67.5°C or 80°C. Native whey protein content decreases with the heat treatment, leading to their aggregation on with the casein micelles. The phenomena depend on the temperature and protein composition. At 80°C, shape ratio Rg/Rh of casein micelles increase with the temperature when the formulation is enriched in lactoferrin. The Rg and Rh decrease when the formulation is enriched in α-lactalbumin and lactoferrin. Transmission Electronic Microscopy are correlated with these results. At 80°C, whey protein aggregates are fixed outside the micelle for the first formulation and they are fixed inside the micelles for the second formulation. Whey protein isolate contains few quantities of caseins. The second objective of this study is to identify the presence of trace of caseins in whey protein isolate using 2D AsFlFFF-RP-HPLC. This technique allows to separate proteins by their size and by their polarity. A whey protein standard mix is studied: α-lactalbumin, β-lactoglobulin A and B, BSA, and 3 caseins. A 2D map is obtained with all separated proteins

    Qualitative assessment of adult patients' perception of atopic dermatitis using natural language processing analysis in a cross-sectional study.

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    Introduction Atopic dermatitis (AD) is an incurable, inflammatory skin disease characterized by skin barrier disruption and immune dysregulation. Although AD is considered a childhood disease, adult onset is possible, presenting with daily sleep disturbance and functional impairment associated with itch, neuropsychiatric issues (anxiety and depression), and reduced health-related quality of life. Although such aspects of adult AD disease burden have been measured through standardized assessments and based on population-level data, the understanding of the disease experienced at the patient level remains poor. This text-mining study assessed the impact of AD on the lives of adult patients as described from an experiential perspective. Methods Natural language processing (NLP) was applied to qualitative patient response data from two large-scale international cross-sectional surveys conducted in the USA and countries outside of the USA (non-USA; Canada, France, Germany, Italy, Spain, and the UK). Descriptive analysis was conducted on patient responses to an open-ended question on how they felt about their AD and how the disease affected their life. Character length, word count, and stop word (common words) count were evaluated; centrality analysis identified concepts that were most strongly interlinked. Results Patients with AD in all countries were most frequently impacted by itch, pain, and embarrassment across all levels of disease severity. Patients with moderate-to-severe AD were more likely than patients with mild AD to describe sleep disturbances, fatigue, and feelings of depression, anxiety, and a lack of hope that were directly associated with AD. Centrality analysis revealed sleep disturbance was strongly linked with itch. Collectively, these concepts revealed that patients with AD are impacted by both physical and emotional burdens that are intricately connected. Conclusions Qualitative data from NLP, being more patient-centric than data from clinical standardized measures, provide a more comprehensive view of the burden of AD to inform disease management

    Broadcasting human voice to piglets (Sus Scrofa domestica) modifies their behavioural reaction to human presence in the home pen and in arena tests

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    The human-pig relationship develops through visual, tactile, olfactory, and auditory interactions. Our objective was to determine the effect of human voices on the development of the human-pig relationship. We hypothesised that human voice facilitates the development of the human-pig relationship. We studied the behaviour of 90 weaned female piglets divided into three treatments: human presence with (HPV) or without (HP) voice, and control (CTRL). For the HPV piglets, the experimenter was present (idle) for 5 min/day in the pen for three weeks and a female voice was broadcast from a speaker. The HP treatment was the same, but a recorded background noise was broadcast instead of the voice. For the CTRL piglets, only routine husbandry care was provided. Piglets were then tested twice in a 3x3 m test area alone for 5 min and then in the presence of the experimenter for 5 min. For test 1, the human voice was broadcast for HPV piglets and the background noise was broadcast for the others. In test 1, HPV and HP piglets investigated the experimenter earlier and more often than CTRL piglets (P < 0.05). HPV piglets moved sooner in the pen than CTRL piglets (P < 0.05). For test 2, only the background noise was broadcast; HPV piglets were thus deprived of the human voice they were used to. In test 2, HPV piglets expressed more stress reactions: their latency to move was longer compared to the others (P < 0.05). HPV piglets also had more physical and vocal interactions: they stayed in the experimenter area longer than HP and CTRL piglets (P < 0.05), and grunted more (P < 0.05). Finally we measured the time taken to move the pigs from their home pen to the truck to move to the farrowing building; we found no effect of the treatment (P < 0.05). In conclusion, broadcasting a human voice did not modify the pig response to human presence and handling in auditory conditions similar to the interaction sessions (i.e. test 1 and moving). However, not broadcasting human voice (test 2) induced stress responses and increased interactive behaviour, which suggests that piglets identified human voice as part of the experimenter’s necessary properties

    Reduced representation characterization of genetic and epigenetic differentiation to oil pollution in the foundation plant Spartina alterniflora

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    Theory predicts that environmental challenges can shape the composition of populations, which is manifest at the molecular level. Previously, we demonstrated that oil pollution affected gene expression patterns and altered genetic variation in natural populations of the foundation salt marsh grass, Spartina alterniflora. Here, we used a reduced representation bisulfite sequencing approach, epigenotyping by sequencing (epiGBS), to examine relationships among DNA sequuence, DNA methylation, gene expression, and exposure to oil pollution. We documented genetic and methylation differentiation between oil-exposed and unexposed populations, suggesting that the Deepwater Horizon oil spill may have selected on genetic variation, and either selected on epigenetic variation or induced particular epigenotypes and expression patterns in exposed compared to unexposed populations. In support of the potential for differential response to the Deepwater Horizon oil spill, we demonstrate genotypic differences in response to oil under controlled conditions. Overall, these findings demonstrate genetic variation, epigenetic variation and gene expression are correlated to exposure to oil pollution, which may all contribute to the response to environmental stress.

    From refugia to contact: Pine processionary moth hybrid zone in a complex biogeographic setting

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    Contact zones occur at the crossroad between specific dispersal routes and are facilitated by biogeographic discontinuities. Here, we focused on two Lepidoptera sister species that come in contact near the Turkish Straits System (TSS). We aimed to infer their phylogeographic histories in the Eastern Mediterranean and finely analyze their co-occurrence and hybridization patterns in this biogeographic context. We used molecular mitochondrial and nuclear markers to study 224 individuals from 42 localities. We used discordances between markers and complementary assignment methods to identify and map hybrids and parental individuals. We confirmed the parapatric distribution of Thaumetopoea pityocampa (Lepidoptera: Notodontidae) in the west and Thaumetopoea wilkinsoni in the east and identified a narrow contact zone. We identified several glacial refugia of T. wilkinsoni in southern Turkey with a strong east-west differentiation in this species. Unexpectedly, T. pityocampa crossed the TSS and occur in northern Aegean Turkey and some eastern Greek islands. We found robust evidence of introgression between the two species in a restricted zone in northwestern Turkey, but we did not identify any F-1 individuals. The identified hybrid zone was mostly bimodal. The distributions and genetic patterns of the studied species were strongly influenced both by the Quaternary climatic oscillations and the complex geological history of the Aegean region. T. pityocampa and T. wilkinsoni survived the last glacial maximum in disjoint refugia and met in western Turkey at the edge of the recolonization routes. Expanding population of T. wilkinsoni constrained T. pityocampa to the western Turkish shore. Additionally, we found evidence of recurrent introgression by T. wilkinsoni males in several T. pityocampa populations. Our results suggest that some prezygotic isolation mechanisms, such as differences in timing of the adult emergences, might be a driver of the isolation between the sister species

    Limited thermal plasticity and geographical divergence in the ovipositor of Drosophila suzukii

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    Phenotypic plasticity has been repeatedly suggested to facilitate adaptation to new environmental conditions, as in invasions. Here, we investigate this possibility by focusing on the worldwide invasion of Drosophila suzukii: an invasive species that has rapidly colonized all continents over the last decade. This species is characterized by a highly developed ovipositor, allowing females to lay eggs through the skin of ripe fruits. Using a novel approach based on the combined use of scanning electron microscopy and photogrammetry, we quantified the ovipositor size and three-dimensional shape, contrasting invasive and native populations raised at three different developmental temperatures. We found a small but significant effect of temperature and geographical origin on the ovipositor shape, showing the occurrence of both geographical differentiation and plasticity to temperature. The shape reaction norms are in turn strikingly similar among populations, suggesting very little difference in shape plasticity among invasive and native populations, and therefore rejecting the hypothesis of a particular role for the plasticity of the ovipositor in the invasion success. Overall, the ovipositor shape seems to be a fairly robust trait, indicative of stabilizing selection. The large performance spectrum rather than the flexibility of the ovipositor would thus contribute to the success of D. suzukii worldwide invasion

    Valorisation des protéines par maîtrise des fonctionnalités

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    La naturalité des produits alimentaires représente une demande forte des consommateurs des pays développés. La naturalité s’entend comme une réduction de l’emploi des additifs pour la fabrication, le stockage, la distribution ou l’usage des produits alimentaires, mais aussi la préservation des propriétés organoleptique et santé des macro- et micronutriments constitutifs. Les protéines sont des constituants essentiels pour texturer des aliments. Il existe cependant 2 limites à l’utilisation des protéines pour leurs propriétés de texture : une trop faible concentration en protéines ne permet pas de texturer une matrice alimentaire de façon satisfaisante, tandis qu’une trop forte concentration en protéines thermo-sensibles, comme les protéines de lactosérum, conduit à une agrégation excessive ou une gélification pendant leur traitement thermique. Un recours à l’addition d’agents gélifiants ou stabilisants est donc nécessaire pour atteindre la texture souhaitée (solide, visqueuse, fluide). Dans le cadre d’un projet interrégional, Profil, nous avons étudié le rôle de différentes fractions protéiques du lait pour contrôler la texture des émulsions sur une large gamme de concentration en protéines de lactosérum et sans ajout d’additif en combinant les propriétés de stabilité thermique des assemblages de protéines de lactosérum et les propriétés interfaciales des caséines. A teneur faible en protéines, notamment en caséines, les assemblages de protéines de lactosérum s’adsorbent à l’interface et connectent les globules gras, ce qui permet de texturer l’émulsion. A plus forte concentration en caséines, celles-ci couvrent l’interface au détriment des assemblages de protéines de lactosérum qui restent dans la phase dispersée et les émulsions sont fluides et stables après traitement thermique, même à forte concentration en protéines. Ces travaux ouvrent une voie innovante pour la création de produits laitiers sans additif répondant à la demande de naturalité des consommateurs. La compréhension du rôle de l’interface eau/huile dans la texturation des émulsions et son ingénierie constituent des leviers pour mieux contrôler la texture de produits existants. Ils offrent également des pistes pour la création de nouveaux produits correspondant à de nouveaux usages et de nouvelles habitudes de consommation. Les assemblages protéiques façonnés par traitements thermomécaniques permettent d’obtenir des propriétés fonctionnelles adaptées à un usage culinaire cible. Or, ces traitements impactent la structure des protéines et partie de leurs propriétés santé. Pour pleinement répondre aux enjeux de naturalité des produits alimentaires, la préservation des propriétés santé des protéines constitue également un enjeu majeur de l’alimentation de demain. Des pistes sont actuellement à l’étude pour répondre à ce double enjeu de maitrise de la techno-fonctionnalité et de la préservation de la bio-fonctionnalités des protéines

    Mycoplasma pneumoniae infections, 11 countries in Europe and Israel, 2011 to 2016

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    Background: Mycoplasma pneumoniae is a leading cause of community-acquired pneumonia, with large epidemics previously described to occur every 4 to 7 years. Aim: To better understand the diagnostic methods used to detect M. pneumoniae; to better understand M. pneumoniae testing and surveillance in use; to identify epidemics; to determine detection number per age group, age demographics for positive detections, concurrence of epidemics and annual peaks across geographical areas; and to determine the effect of geographical location on the timing of epidemics. Methods: A questionnaire was sent in May 2016 to Mycoplasma experts with national or regional responsibility within the ESCMID Study Group for Mycoplasma and Chlamydia Infections in 17 countries across Europe and Israel, retrospectively requesting details on M. pneumoniae-positive samples from January 2011 to April 2016. The Moving Epidemic Method was used to determine epidemic periods and effect of country latitude across the countries for the five periods under investigation. Results: Representatives from 12 countries provided data on M. pneumoniae infections, accounting for 95,666 positive samples. Two laboratories initiated routine macrolide resistance testing since 2013. Between 2011 and 2016, three epidemics were identified: 2011/12, 2014/15 and 2015/16. The distribution of patient ages for M. pneumoniae-positive samples showed three patterns. During epidemic years, an association between country latitude and calendar week when epidemic periods began was noted. Conclusions: An association between epidemics and latitude was observed. Differences were noted in the age distribution of positive cases and detection methods used and practice. A lack of macrolide resistance monitoring was noted

    High-resolution assessment of French grassland dry matter and nitrogen yields

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    Grasslands offer many environmental and economic advantages that put them at the heart of future sustainable ruminant production systems. This study aimed to quantify and map the dry matter yield (DMY) and nitrogen yield (NY) of French grasslands resulting from cutting and grazing practices, based on the existing diversity of grassland vegetation, management, soil and climate conditions, using a research version of the STICS crop model called PâturSTICS. This model simulates daily dry matter (DM), nitrogen (N) and water fluxes involved in the functioning of grasslands and crops in response to management and environmental conditions. It was improved to represent deposition of animal waste on grassland soils during grazing and to simulate DM production and N content of grasses and legumes more accurately. Simulations were performed for locations across France on a highresolution grid composed of pedoclimatic units (PCU) obtained by combining the spatial resolutions of climate and soil. The main grassland types and associated management types were determined for each PCU and then simulated over 30 years (1984-2013). Using the simulated values, predictive metamodels of annual grassland DMY and NY were developed from easily accessible explanatory variables using a random forest approach. Annual model predictions were aggregated and averaged at the PCU scale, then compared to regional observations. Predicted DMY agreed with available observations, except in semi-mountainous and mountainous regions, where PâturSTICS tended to overpredict DMY, probably because it ignores effects of snow, frost and slope, and due to how it represents effects of temperature and water stress on plant growth. According to results, three-quarters of French grasslands produce and export at least 7.6 t DM ha-1 yr-1 and 172 kg N ha-1 yr-1, respectively. One-quarter of French grasslands produce and export at least 10.7 t DM ha-1 yr-1 and 254 kg N ha-1 yr-1, respectively. The latter are located mainly in northwestern France, the north-western Massif Central, the French Alps and the western Pyrénées, all of which have environmental conditions favourable for grass growth. The metamodels developed are interesting proxies for PâturSTICS' predictions of grassland DMY and NY. Our results provided valuable knowledge that promotes better use of the potential forage production of French and European grasslands to improve protein selfsufficiency and N fertilisation management in ruminant livestock systems

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