Ferchaud Stéphane

Virginie Lefoul

Variability of piglets' adaptability at weaning in organic farming system

Keywords: weaning, piglet, adaptation, heterogeneity, behaviour

In organic farming, weaning piglets remains a critical period, although later than in the conventional system. The use of hormones being forbidden in organic system, heat synchronization is absent, which leads to spread of birth associated with heterogeneous ages, weight and health status of the piglets. The differences can be accentuated by a poorly adapted solid diet before and after weaning. The aim of the study was to understand the difference involved in the physiological mechanisms in piglet adaptation strategies to propose adapted feeding strategies. The studied population of fifty piglets, from four genetic lines, comes from 5 sows (Landrace x Duroc) of the INRA-Genesi unit and a boar (Pietrain x Large White). Five batches, of ten pigs per sow, were constituted by weight range: four "large" piglets, two "medium" and four "small". They had access to a second-age feed from ten days and to roughage, high protein, after weaning (Wrapped alfalfa). Animals weights were recorded on days (d) 1-10-21-28 42-48-55-62. Blood samples were collected at d 42-48-55-62 for inflammatory indicators and oxidative stress measurements around weaning. Behavioural observations in farrowing room at d 15-20-30-40 and at post-weaning at d 43, were carried out to establish weaning adaptation profiles. The results showed that the small piglets at d 42 were on the posterior teats and that there was no clear indication if the piglets ate more solid feed in the farrowing room compared to the larger piglets. At post-weaning, it was shown that the small piglets were on average more present in the area of feeding interest than the larger ones. The smaller piglets also had a higher haptoglobin rate than the large and medium piglets. Finally, there was no weight compensation from the smaller piglets compared to the mediums and larger piglets. The average weight significantly differs between all three categories at d 42 and d70.

No part of this publication may be reproduced or published in any form or by any means, electronic, mechanical, photocopying, recording or otherwise, without prior written permission of the head of the Adaptation Physiology Group of Wageningen University & Research, The Netherlands Niets uit dit verslag mag worden verveelvoudigd en/of openbaar gemaakt door middel van druk, fotokopie, microfilm of welke andere wijze ook, zonder voorafgaande schriftelijke toestemming van de hoogleraar van de leerstoelgroep Adaptatiefysiologie van Wageningen University & Research. Table 1:

LIST OF FIGURES

LIST OF TABLES

INTRODUCTION

In organic pig farming, the weaning period remains, as in conventional farming, a critical period despite the later age of weaning (6-8 weeks vs 3-4 weeks in conventional farming) implying a greater digestive, behavioural and immune maturity of piglets. At weaning, piglets face abrupt changes in their social and physical environment, such as break-up of the motheryoung bond, change of feeding behaviour, reunion with unfamiliar animals from different age, weight, sanitary status, with negative social interactions and change of environment (Beattie,2000;[START_REF] Campbell | The biological stress of early weaned piglets[END_REF]. Although the adaptation dynamics of piglets after weaning in conventional farming are well known, piglets adaptation weaned at 42 days in organic farming are less studied and therefore are much rarer.

In the present study, we mainly focalized on organic farming systems, in which the lack of synchronization of estrus causes spread births over time. These factors lead to a high heterogeneity in age, weight and thus in maturity of piglets at weaning. The heterogeneity can be accentuated by a creep feed and pre-starter diet that are still poorly adapted to young piglets because of constraints related to the organic farming specifications such as the prohibitions of use of synthetic amino acids or lack of milk proteins from organic grade. The feed distributed before and after weaning has a high protein content and is not always well balanced and digestible for piglets [START_REF] Mavromichalis | Pig Nutrition, weaning & Farrowing[END_REF]. It was found that piglets may consume creep feed in compensation for insufficient milk production by the sow (Barber et al., 1955;[START_REF] Algers | Behaviour and weight changes at weaning and regrouping of pigs in relation to teat quality[END_REF]. Preliminary results, however, have shown that the disadvantages of this heterogeneity can be overcame by the ability of smallest piglets to express compensatory growth in the presence of roughage silage with high protein content (wrapped alfalfa for example), suggesting that such a behavior is one of the adaptive strategies of small piglets to weaning (ITAB, 2014).

The aims of the study are to characterize the adaptation strategies within a group of heterogeneous piglets and to understand the behavioural and physiological mechanisms involved in the adaptive strategies. This means that individual within a heterogeneous group could benefit from this variability in term of management and practices as, for example, adapted feeding strategies. For that purpose, we recorded behavioural, physiological and performance traits at an individual level in an "organic-like" system in order to identify profiles of piglets with diverse adaptive strategies to weaning. The project was divided in two parts: observations and analyses during the lactating period in order to characterize the "early status" of the piglets that will be observed and analyzed during post weaning period.

More specifically, during the lactating period, our hypotheses were that the small piglets at 42 days (d 42) were also small at d 1, then that the small piglets suckled the posterior teats of the sow, and at last that the small piglets would eat solid feed available at d 10 while the larger pigs would continue drinking sow's breastmilk. During the post weaning period, we hypothesized that the smaller, slower-growing piglets would be expected to eat more roughage silage with high protein content, then still the small piglets would have a higher inflammatory state compared to the larger piglets at weaning and at last that the small piglets would have compensated their low body weight by catching up the larger piglets maybe by eating wrapped alfalfa available at weaning.

Our experimental strategy was to measure traits related to growth (body weight), behaviour, health and underlying physiological mechanisms such as microbiota, inflammation, immunity, metabolism in 50 piglets from five litters. This report presents only the results of growth, behaviour, as well as blood indicators of oxidative status and inflammation. To enhance the variability, piglets were produced from four genetic types: one Large White x Pietrain boar and five Landrace x Duroc sows. The measured traits were then used to establish classes of pigs with different strategies or patterns of adaptation / nonadaptation to weaning and to explore the biological mechanisms that characterize these classes of animals.

LITERATURE REVIEW

At weaning, piglets face abrupt changes in their social and physical environment, such as break-up of the mother-young bond, change of diet, reunion with unfamiliar animals from different age, weight, sanitary status, with negative social interactions and change of environment (Beattie,2000;[START_REF] Campbell | The biological stress of early weaned piglets[END_REF]. Today, in Europe, piglets are usually weaned between 21 and 28 days of age. The Council directive says: "No piglets should be separated from their mothers until they are 28 days old, unless the welfare or the health of the sow or piglet requires them to be separated sooner. However, piglets can be weaned up to seven days earlier if they are moved to specialized premises which will be emptied, cleaned and disinfected completely before the introduction of a new group and separated from the premises where the sows are kept to minimize the risk of disease transmission to piglets". In France the average age of weaning is 23.9 days related to increased number of litters per year for a sow and expecting reduced risk transmission from sows to piglets. However, the biological characteristics of swine describe weaning occurring later at around 10 to 12 weeks of age following gradual physiological and behavioural processes [START_REF] Orgeur | Mother-young relationships in pigs: from birth to weaning[END_REF].

This literature review explains at first the weaning conditions of piglets and what it implies as adaptation. Then in a second part the major issues of weaning process is described before a description of weaned piglets in organic farming in comparison with the conventional farming will be point out.

Weaning of piglets and what it implies as adaptations

The newly weaned piglet has to adapt itself to a total new situation in the absence of the mother, with a new diet, with a transition from a milky diet to a solid diet, a new housing condition, and in which it is often met with new conspecific by formation of homogeneous weight groups. The weaned piglets are also subjected to a new microbism with the grouping of animals of different origins (different litters). Weaning is then defined by all these conditions reunited.

Change of Feeding

During the two first weeks of lactation, piglets get at the same time their nutrition needs (water and feed) via the sows' milk; later on, piglets must distinguish thirst from hunger and learn how to satisfy theses needs via separated media as milk, feeder and water. Therefore, animals must adapt themselves to new conditions through behavioural and physiological adaptations [START_REF] Campbell | The biological stress of early weaned piglets[END_REF]. Studies have shown that piglets only drink milk in their first two weeks of life and from two weeks they diversify their feed. Moreover, from three to four weeks of age, studies have shown that the milk production of the sow is no longer sufficient to cover the needs of piglets, or at least of all piglets with a large litter. [START_REF] Jones | Rick Jones-Farrowing and Lactation in the Sow and Gilt. The pig site[END_REF]). However, after weaning, piglets are generally fed with a feed "phase 1 or pre-starter" called starter which is distributed in a dry granules form and contains proteins from milk origin such as milk powder, lactoserum or lactose (IFIP, 2007). While sow milk is composed of 20% dry the "pre-starter diet" contains about 88% dry matter. The nutritive composition of milk differs from that of pre-starter diet. Indeed, the maternal milk dry matter is composed of 30% protein, 40% fat and 25% of lactose compared to the "pre-starter" feed in which the dry matter is composed of about 18 to 20% protein, 6 to 7% fat and complex sugars under starch form (plant orgin).

Maternal milk is also a highly digestible and an important immune sources feed for piglets [START_REF] Williams | Growth of the weaned pig[END_REF]. Under the mother, the piglet suckles a little amount of milk up to 24 times a day. It provides the piglet immunoglobulin A which provides passive local immunity and immunoglobulin G which provides systemic immunity. It also provides non-specific antiinfectious components such as lactoferrin or lysozyme [START_REF] Bernard | Le lait de truie: composantes humorales et cellulaires de la protection pour le jeune porcelet[END_REF]. Plus, a piglet has its own teat around five days old. A study have shown that there is no influence on teats' order concerning sex, weight at birth and birth order, however the piglets that are positioned on the four front teats pairs have a tendency to gain a little more weight than the others [START_REF] Rossilon-Wanier | Development and consequences of teat-order in piglets[END_REF].

At weaning time, the piglet receives directly a new food less digestible and less appetizing, largely composed of proteins of plant origin (as soy, cereals), to which it must adapt without delay because of the few body reserves available to him. Indeed, the piglet goes to the trough when the lights are on and eat less during the first days of weaning [START_REF] Bruininx | Individually measured feed intake characteristics and growth performance of group-housed weanling pigs: effects of sex, initial body weight, and body weight distribution within groups[END_REF]. Thus, to reach the same energy level as pre-weaning, the piglet should ingest larger amounts of food. However, the piglet's digestive tract is not yet well developed to accept a large quantity or the quality of this post-weaning food less digestible [START_REF] Williams | Growth of the weaned pig[END_REF]. Regarding the inflammation due to less immune sources, recent studies have shown an increase in the expression of genes coding for pro-inflammatory cytokines and an infiltration of inflammatory cells in the digestive mucosa during the first two days after weaning [START_REF] Lalles | Gut function and dysfunction in young pigs: physiology[END_REF]. This switch from milk feed to solid feed leads also to behavioural changes. Indeed, before weaning, feeding behaviour is part of a breastfeeding sequence between the sow and the litter, in which obtaining milk depends on the auditory exchanges (grunts of the sow) and tactile (massages of the udder and suction of the nipple by the piglets). After weaning, the feeding behaviour is associated with the ingestion of dry food, and is expressed by the movement of chewing (Meunier-Salaun and Picard, 1996).

Change of housing, environment and mixt with conspecifics

Piglets face abrupt changes in their social and physical environment at weaning, reunion with unfamiliar animals from different age, weight, sanitary status, with negative social interactions and change of environment (Beattie,2000;[START_REF] Campbell | The biological stress of early weaned piglets[END_REF]. Piglets are moved into a new building which could be on the same site or in another farm. At their arrival, the piglets are grouped together, according to their weight and sex, in order to create uniform weight groups of animals. The mixture of individuals of different litters leads to agonistic behaviours associated with the establishment of a hierarchy of dominance and subordination [START_REF] Fels | Influence of origin litter on social rank, agonistic behaviour and growth performance of piglets after weaning[END_REF]. These indeed, limited-timed behaviours lead to injuries, which are generally not serious as fighting behaviour disappears after 24 to 48 hours, to give way to threat or avoidance type of behaviour [START_REF] Friend | Behavior and performance of pigs grouped by three different methods at weaning[END_REF]. Nevertheless, these injuries can be a potential source of inflammation and a gateway for germs. In addition, during the first week of post-weaning, the piglet has an energy deficit due to its diet. This deficit can be accentuated by the energy spent during fighting for hierarchy, or fighting against the cold in an environment not suitable for young piglets.

In the weaning process, the underlying social mechanisms are a mother-young bond that becomes more and more distended as time goes by, and in parallel socialization within the litter will orientate the piglet towards an environment far from the mother and thus brings the piglet to a social autonomy in regards to the mother. In addition, socialization can be done either within reach, or with other litters before weaning if the litters have the opportunity to meet, such as outdoor system or maternity systems building allowing a mixture of litters before weaning. This socialization before weaning is a way of minimizing stress at weaning by preparing the animal for a larger social environment and thereby improving performances. [START_REF] Hessel | Socializing piglets before weaning: effects on behavior of lactating sows, pre-and postweaning behavior, and performance of piglets[END_REF] To sum up, weaning is a sensitive period in a piglet's life as it is a source of stresses. The consequences of post-weaning can be expressed under physiological and behavioural form of disturbances.

Welfare and health matters of weaned piglets

Welfare and Health are two notions often discuss in pig farming. Pig production is considered as a production is which the level of animal welfare is quite low due especially to a poor environment (small surface, poor enrichment and rooting materials) and the reunion with unfamiliar piglets. The poor environment limits the piglets to express their natural beahviour as nosing, chewing and rooting which could lead to abnormal behaviour as belly noosing on the unfamiliar piglets during the reunion with different litters. There are new regulations concerning animal welfare in which space are widen and rooting materials is necessary and there is also a new regulation regarding the use of antibiotics.

Animal Welfare

The pig welfare is assured by Council Directive 2008/120/EC which lays down the minimum standards for pig protection in Europe. More and more the Council Directive are working to improve the quality of pig's welfare by: improving flooring surfaces, increasing the living space available for sows and gilts, setting requirement for light and noise level, setting a minimum weaning age of four weeks, and providing permanent access to fresh water and enrichment materials for rooting and playing (FAO, 2008).

One of the newest regulations concerned the pregnant sows that must be kept in groups instead of individuals stall during their pregnancy. Indeed, apart from farrowing period and boars, all pigs are to be raised in groups and must be provided access to water and food of appropriate quality at regular intervals for all the animals at the same time. Plus all pigs must have access to a sufficient quantity of enrichment materials that does not compromises their health and which could avoid abnormal behaviour and in contrary enable the pigs to carry out their proper investigations activities and fulfil their natural behavioural needs. Moreover, tail docking and teeth resurrection as a routine are forbidden, the Commission has developed several activities to prevent cannibalistic behaviour. Tail is an intervention during which a portion of the tail of the piglets is definitively sectioned in its first week of age. The cut of the tail is performed to prevent or reduce tail bites. The bite of the tail is an important symptom that comes from many factors, such as lack of space to express natural behaviors, inadequate building atmosphere, and poor quality food or health problems. A factor very important risk is the absence of rooting materials (straw or litter type) for animals to express their natural behaviour of noosing. In many intensive pig farming systems, animals have very limited opportunities to search with their snouts or seek food. Pigs are very curious and intelligent. When they have nothing else to do, they begin to explore and play with the tails of their peers. Some chew the tails of their peers causing injury. Then, teeth resurrection is an operation that involves cutting (trimming) or grinding the upper part of the corner teeth of young piglets. Teeth resurrection is done to prevent injury between piglets from the litter and on the sows, when the piglets are fighting for the best teat on the sow's udder. For this reason, all inadequate environment conditions and stocking density management systems must be changed (FAO, 2008).

Reduction of antibiotic

Antibiotics are widely given to humans and animals, both as curative and preventive drugs. In any case, their use is likely to lead to the emergence of bacterial resistance in humans and animals through mechanisms channeled by the environment or food. Fifty percent of the antibiotics produced in the world are used for pets and farm animals (Le Foll, 2017).

The project "Ecoantibio" is a multi-year plan put in place by the Agriculture Ministry in France and is part of a wider project known as the Agro-Ecology Project. It relates to all animal sectors and covers all French territories. Numerous exchanges with representatives of agricultural and veterinary professional organizations, scientists and representatives of veterinary pharmaceutical laboratories identify the proposals for actions of the national plan for reducing the risks of antimicrobial resistance in veterinary medicine for the period 2012-2017. This plan is consistent with the 2011-2016 national antibiotic alert plan conducted by the ministry in charge of health as well as the European Commission's action plan to fight against threats of antimicrobial resistance. The plan "Ecoantibio" 2012-2017 planned a 25% decrease in the use of antibiotics in five years. Prescription and antibiotic use practices have already evolved considerably as stopping the use of antibiotics as a growth factor in food in 2006, and these practices are still evolving.

Despite the implementation of these measures, the weaning period is the physiological stage in which the consumption of antibiotics remains the largest in pig production. Indeed, 57% of the antibiotic treatments applied during a farm pig's life are administered during the post-weaning period, mainly as a result of, or in the prevention of digestive disorders (see Review Hemonic, 2014). Therefore, the weaning period is the subject of a lot of attention and concentrates the efforts of research on alternative solutions allowing the preservation of the health of the weaned piglets and therefore technical performances.

Weaned piglets in organic farming vs conventional farming

Conventional farming results from the transformations initiated in the fifty's in Europe and is characterized by the use of high-yielding varieties and breeds, by inputs that make it possible to optimize production (fertilizers and pesticides or concentrated foods, and strong medication) on highly specialized and mechanized farms. In contrast to conventional agriculture, organic farming excludes chemical inputs and any systematic use of drugs, per consequence excluding "intensive" breeds. It has become today more a "specification" than an agricultural model [START_REF] Vogels | Agriculture biologique, agriculture conventionnelle : explications[END_REF].

Organic pig production meets strict regulation specifications that evolve regularly to meet the demand of consumers, breeders, but also the industry. It is a niche production. It is a farming system environmentally friendly and sustainable. In 2015, there is more than 960 000 certified organic pigs in the European Union, which represents only 0.6% of the herds. The main breeders are Denmark, France, Germany and the Netherlands. The share of organic certified flocks remains very low (maximum 1.7% in Denmark, 0.9% in France and 0.6% in the Netherlands) (Agence Bio, 2016).

The principle of weaning in pig farming is the same in all breeding systems. However, the main difference between a "conventional" ( and "organic" system is a matter of timing, the physiological and behavioral processes involved being accelerated in the conventional system, hence the notion of "early weaning" in "conventional" and "late weaning" in the organic system. Weaning in early weaning conditions is in fact associated with other authorized interventions (prohibited in organic farming) such as teeth resurrection, tail docking, iron injection and castration done during their first week of life, which add high stress factors for piglets in conventional system during weaning that induces changes of feed and social behaviour. The administration of iron is done due to little iron reserve which is scarce in sow's milk. It is performed to avoid a drop in hemoglobin which can lead to anemia. Organic piglets must find a source of iron elsewhere, for example in the soil and dirt. Regarding teeth resurrection and tail docking it is performed in conventional system to reduce wound to conspecific at weaning. Forbidden in organic farming, it is essential to find alternative solutions to avoid cannibalistic behaviours between the animals. For this reason, it is necessary to add enrichment material in organic weaned piglets' environment and widen the surface per piglet.

In organic farming, housing conditions for animals must meet their physiological and ethological needs (including behavioural needs freedom of movement and comfort). Outdoor spaces must offer protection sufficient against rain, sun, wind and extreme temperatures (Chambres d'Agriculture de Bretagne, 2011). There are several objectives of an organic pig farming concerning the feeding condition. First, feed must mainly be produced on the farm or come from organic farms from the same region. Then roughage, fresh, dry or ensiled should be added to the daily ration of pigs. Piglet's feeding is based on natural milk, preferably maternal, for 40 days minimum (vs 21 days in conventional system). Moreover, the use of GMO feeds, antibiotics, coccidiostats, drug substances, stimulants growth or any other substance intended to stimulate growth or production of piglets are prohibited in organic farming (versus authorized in conventional system) (Chambres d'Agriculture de Bretagne, 2011). This also means that the administration of hormones in order to synchronize the birth of all the piglets from the same group of sows to have homogeneous piglets at weaning age is forbidden in organic farming. This condition is one of the major problem encounters in organic farming as it could have economic impact for the farmer and performances impact for the piglets. Indeed, the absence of synchronization of estrus causes spread births over time. These factors lead to a high heterogeneity in age, weight, maturity and health status of animals at weaning period. The heterogeneity can be accentuated by a creep feed and prestarter diet that are still poorly adapted to young piglets because of constraints related to the organic farming specifications such as the prohibitions of use of synthetic amino acids or lack of milk proteins. Heterogeneity of piglets at weaning could be an economic lost for the farmer as he sells his pigs per group weight. The smaller, thinner piglets would cost much less than the larger piglets. Very few studies have been done regarding the consequences and the adaptability of the piglets at weaning in organic farming. Plus nowadays, intensive farming system is being more and more criticized.

It is for this reason that the present study, mainly focalized on organic farming systems, to understand the behavioural and physiological mechanisms involved in heterogeneous piglets' adaptation strategies.

MATERIALS AND METHODS

Animals, Housing and Feeding

The experiment was conducted in the experimental unit of INRA GENESI-Rouillé, in "organic-like" conditions, in compliance with the French directive on animal experimentation and care . The analyses were conducted at INRA GENESI-Rouillé and INRA PEGASE-Saint Gilles. Piglets ((Large White x Pietrain) x (Landrace x Duroc)) from 5 litters were observed during the whole study from birth day to 70 days (d70). At birth, piglets were identified with an ear tag with a unique number inserted on their left ear. During farrowing, the five sows were transferred to one farrowing room one week before the farrowing term day. The farrowing room was equipped with conventional individual pens (1.80 meters x 2.25 meter) with one crate for each sow (0.65 meters x 1.90 meters) with partly slatted floor. The farrowing room was indeed conventional, however only the infrastructure of the farrowing room was different of organic farming rules due to lack of space. Piglets were not castrated, and teeth and tails were kept intact. They were weaned as organic farming at 42 days. The room was forced ventilated and heated at a temperature of 26°C and gradually decreased at 24°C at 3 days after farrowing. Each pen had carpets 80 cm long and 40 cm large and two infrared-heated lamps, on both side of the sow. At d 3 one infrared-heated lamp per pen was removed. At d 10, Phase 2 feed supplying 17.7% of crude protein was provided ad libitum in each pen in a small feeder. The sow's lactation feed was distributed twice a day starting with a quantity of 2.4 kg at each distribution and was increased by 300 g every day up to a quantity of 5 kg a day. Then at d 28 each pen was extended of 80 cm long. Straw on the extended part of the pen was added and change every two days for the piglets as enrichment (Surface of straw bedding: 1.44 m 2 . At weaning at d 42, ten piglets were selected in each of the five litters, according to their bodyweight: the four smallest, the two mediums and the four heaviest piglets from each litter were chosen to form five heterogeneous batches to mimic organic conditions. A total of 50 piglets were then allotted in five pens of 10 weaned piglets each in order that each pen was composed by five pairs of piglets from each litter (5*2 =10 piglets). The post weaning pens were a large stable on concrete floor with straw bedding, with outdoor climate. Each pen sized 3.5m x 3.5m was equipped with a kennel with straw litter, one watering place, a feed rack for wrapped alfalfa distribution and a feeder. The feed is a commercial pelleted feed adapted for the second phase of the growing of the weaned piglets from ALICOOP and silage were distributed ad libitum. For the video analysis, Bascom cameras related to a recorder were installed. Due to lack of material, only four cameras were available. Each camera was placed, on top, in the middle of each pen except the last pen due to one camera missing.

Piglets' behaviour during lactating period

The purpose of this observation was to assess the piglets' interest for solid feed. These observations were done twice a day, at 9:00 in the morning and at 3:00 in afternoon at d15, d 20, d 30, and d 40. At each day, three observers were present and recorded two pens each for a total duration of one hour. Within each litter, piglet behaviours were recorded every five minutes. This means that one observer observed their first pen at initial time 0 minute before switching to the second pen at 2:30 minutes and then come back to the first pen at 5:00 minutes. The recorded behaviours (Table 1) were: exploratory (E), pre-suckling and suckling phase (A), drink water (B), conspecific manipulation (C), none-active (I), eat solid feed (M) and others (O). The percentage of the total time observed for each behaviour was then calculated for each pen.

Piglets' behaviour after weaning

After weaning, piglets of each pen were video recorded at d 43 and d 47 for a total duration of 12 hours. However, due to lack of time, only the first day, d 43, and for a total duration of 4 hours was analysed. We have chosen to look up especially a 4 hour-period in which the piglets were the most active (the validation period of 4 hours compared to the 12 hour-period was presented in Appendix 1). One observer performed a continuous observation using the

Behaviours

Letter Descriptions

Pre-suckling and suckling phase

A Piglet near the udder (distance <body length of a piglet): turned towards the udder, active, stimulates the udder or suckle.

Drink

B

Presence at the drinker, contact or drink of water Explore E Piglet is active in the crate, explores the crate, mastic fence or trough.

Conspecific manipulation

C

Piglet nibbles the tail, ears or the body of one of its conspecific. (Tail biting, belly noising…) Eat M Presence of piglet in the trough, head of the animal in the trough with or without consumption, head rose above the trough with chewing movements.

Non-active

I Piglet is standing still, lying down, sitting or kneeling, without major motor activity.

Other O Play (jump, run), defecation, urinate, or other observed behaviours than those described above.

Observer XT software and recorded the presence and the proximity (less than one piglet length) of each piglet at different Zones (Figure 1) defined as followed: Zone 1 the area of the feeder with the feed; zone 2 the watering place; zone 3 the area of the feed rack with wrapped alfalfa distribution; zone 4 the area between the three first zones; zone 5 out of the feeding area. The total duration spent in each zone was automatically calculated by the software.

STATISTICAL ANALYSES

Before the weaning period, observations and analyses were done on all piglets born and alive (n=58 piglets after an age of 48 hours) to characterise the "early status" of the piglets. Fifty piglets were included in the study to ensure proper statistical results. However only Forty piglets were included to the behavioural analysis during post weaning due to one camera missing for the last ten piglets. Litters from five sows were used to compose the post-partum groups in which the four largest, two mediums, the four smallest piglets from each litter were chosen in order to keep and create a maximum of heterogeneity at weaning. All analyses were performed with the statistical R software using R Studio and R Gui, the piglet considered as the experimental unit. The R packages used are FactoMineR and dyplyR.

Animal studied were allotted in the different post weaning pens at 42 days.

The preliminary analyses led us to classify all the piglets of 42 days of ages in three groups according to their Body Weight at d42: Small [5.94; 12.37 kg], Medium [12.43;13.97 kg] and Large [14.05; 18.37 kg] piglets. The average weight of all piglets at d42 is 13.17 kg. These groups served as reference for all observations and analyses during lactating and postweaning periods.

Individual dataset was submitted to multivariate analyses in order to identify the profiles of piglets. In a first step, ANOVA analyses of variance were done to determine differences between small, medium and large piglets regarding the teats position. In order to test our initial hypotheses regarding behaviour, a Principal Component Analyses (PCA) was run followed by a Clustering classification to establish classes of animals with different patterns of adaptation to weaning period. At last, an ANOVA was done to answer the question whether the small piglets had compensated their low BW in order to catch up the large piglets.

A biostatistician (Rémi Resmond) from INRA-PEGASE helped and supported all the analyses.

Poids des Porcelets selon leur classes d'appartenance au Sevrage

Poids J42 en kg

RESULTS

This part is presented in two sections: one is about the observations and analyses done during the lactating period and one about the observations and analyses done during the postweaning period. The initial hypotheses are reminded in each section.

Analysis during the lactation period

The first hypothesis is that the small piglets at d 42 were also characterized as small at d1. The second hypothesis is that the small piglets at d 42 would have been drinking milk from the posterior teats of the sow. The last is that the smaller piglets eat the solid feed available at d 10 while the large piglets would continue drinking the sow's breastmilk.

Results on weight of the piglets at 1 day of age

A state-transition map is represented in Figure 3 and is used for a descriptive analysis of piglet transition between their category of weight at d 1 and their category of weight at d 42.

Twelve piglets that were classified as small piglets at d 42 were already "small" at d 1. One small piglet at d 1 was categorized as Large at d 42 and four were categorized as Medium sized at d 42.

Results on the piglet's teat position on sow

The numbers of piglets from each weight category at d 42, illustrated in Table 2, are present on the three teats position (Anterior teats, Middle teats and Posterior teats). We obtained 11 small piglets on the posterior teats and 11 large piglets on the middle teats. A khi 2 test revealed a significant difference (p-value < 0.0001) between teats position and weight category of piglets at d 42.

Results of the scan sampling observation

The Figures 4 and5 The Figure 5 shows two clear axes for the individual map. Dimension 1 would be the time, for morning on the left at 9:00 and 3:00 in the afternoon on the right. Dimension 2 represents the age with younger age on top and older age on the bottom.

To summarize what we observed from these two figures, we notice that piglets were more inactive in the morning and more active in the afternoon. Moreover, piglets were more present at pre-suckling and suckling phase at younger age and were more present eating solid feed from age d 30.

Figure 6 represents the individuals added on the Factor Analysis of Mixed Data (FAMD) map. Taking into account the same dimensions axes of figure 5.

By adding the individual on the FAMD map, we noticed that there is no clear distinction between the Small, Medium and Large piglets. We could not conclude on whether a piglet characterized as Small at d 42 goes to eat solid feed more than suckling compared to a large piglet.

Analysis during Post-weaning period

The first hypothesis was that the smaller, slower-growing piglets would be expected to eat more creep feed. The second hypothesis is that the smaller piglets would have a higher inflammatory level compared to the large piglets. The third hypothesis was that the smaller piglets would have compensated their low body weight by catching up the larger piglets by adapting themselves.

Figure 6 : Representation of the Factor Analysis of Mixed Data (FAMD) of all piglets during the 8 scan sampling

Weight group:

• Small • Medium • Large

Results of the video observation

The Figure 7 represents the PCA of the variables recorded by video observation at d 43 (the day after weaning). The variables are the total time a piglet spent at eating, being at proximity or laying at the different zone. The axes colors determined the contribution of the variables to the two first axes. Dimension 1 represents the different zones of interest (Figure 7): on the left side of the graph piglets spent a major part of their time in out-zone (Zone 5) that is the non-feeding zone. The right side represents the feeding zone and more precisely once the piglets are eating in Zone 1 (feeder with phase 2 feed) and eating in Zone 3 (Wrapped alfalfa rack). Dimension 2 isn't really clear enough to conclude.

The Figure 8 represents the individual placed on the variable PCA analyzed on Figure 7. We notice that the small piglets spent a major part of their time eating at the feeder, the medium piglets were eating the wrapped alfalfa feed at the rack, and that the large piglets spent most the time in Zone 5 that is out of the feeding areas.

To summarize, the small piglets tend to spend more time in the areas of interest (zones 1, 2 and 3) compared to the large piglets with a pvalue = 0.063. The average time was calculated as the total time spent in each zone divided by the number of visits.

Blood analysis

Four blood samples were collected at d 42 in the farrowing room and at d 48, d 55, d 62 in the post weaning building. There is an impact of weaning with significant greater haptoglobin plasma concentrations on d 48 compared to d 42, d 55 and d 62 (<0.001). Plus, there is also an impact of weaning with significant greater oxidative products (d-ROM) on d 48 compared to d 42 (<0.001) (Figure 9; Appendix 2). The unit d-rom is CARRU named Carratelli unit: one CARRU 0,08 mg H2O2/100mL

The small piglets had higher haptoglobin plasma concentrations than Medium and Large piglets (< 0.05) and no difference for BAP (total antioxidant status measured in the plasma) between each category of weight. The small piglets had greater oxidative products (dROM) than the Large and Medium piglets.

DISCUSSION

The aim of the study was to characterize the physiological mechanisms in piglet adaptation strategies with the ultimate goal to propose adapted feeding strategies for smaller piglets to compensate their low body weight in organic farming.

Lactation period

The first hypothesis was that the piglets categorized as small piglets at d 42 were also categorized as small at d 1 and as a consequence did not compensate their weight during the lactation period. A study by [START_REF] Gondret | Influence of piglet birth weight on postnatal growth performance, tissue lipogenic capacity and muscle histological traits at market weight[END_REF] showed that piglets born light had a lower average daily gain during suckling and post-weaning periods. However, our results showed that five piglets changed to a larger category from d 1 to d 42. Twelves piglets were already characterized as small at d 1. Indeed, one small piglet at d 1 was categorized as Large at d 42 and 4 were categorized as Medium at d 42.These 5 piglets did compensate somehow during the lactation period. One explanation of the discrepancy between the present study and [START_REF] Gondret | Influence of piglet birth weight on postnatal growth performance, tissue lipogenic capacity and muscle histological traits at market weight[END_REF] could be that the last authors reported the average daily gain compared to this present study in which we looked at a change of category of individual.

According to [START_REF] Rossilon-Wanier | Development and consequences of teat-order in piglets[END_REF], there is no influence on teat order concerning sex, weight at birth and birth order; however the piglets that are positioned on the four front teat pairs have a tendency to gain a little more weight than the others. Our second hypothesis was that the small piglets suckled the posterior teats which could have an influence on their growth rate. Our results showed a significant difference between the piglet's position at the teats and the weight category of piglets at d 42 in which 11 Small piglets were positioned on the posterior teats and 11 Large piglets were positioned on the middle teats. We therefore supposed that the best teats, those producing more milk, could be the one in the middle and not on the anterior teats as expected. Nevertheless, the notion of "best" teat must be taken carefully. Indeed, the production of milk in the sow's teats depends on the sucking strength of the piglets present. The large piglets could be considered as stronger and therefore performed massage stimuli, and stronger suckle on the middle teats of the sow's udder and per consequence more milk would come out from these teats. Another study conducted on 76 litters of Large White x Landrace pigs (Frazer et al., 2009) showed by a regression analysis within litters a significant relationship between suckling position and weight at birth and at 3 weeks of age with the large piglets suckling the anterior teats. However, the correlation coefficient of the regression was quite small. Indeed, the correlation between birth weight and weight at 3 weeks (d 21) was not significant and a difference of more than 0.25 kg in birth weight persisted at d 21 regardless the piglets' position on the anterior teat. The relationship between suckling position and weight at birth or at d 42 is still weak. The last hypothesis was that the small piglets would eat more solid feed available at d 10 while the larger pigs would continue drinking sow's breastmilk. Our results did not confirm this hypothesis and showed that all piglets regardless their body weight went as well to the sow's udder than to eating solid feed. This finding is corroborating by the other results. Indeed, we notice that piglets were more inactive in the morning and more active in the afternoon. This finding confirms the study by [START_REF] Guoan | Effect of Lactation Environment on Behaviour of Suckling Piglets[END_REF] in which the piglets were more lying and walked less in the morning compared to the afternoon where all other activities increased significantly. Moreover, our study points out that the piglets would be more present at pre-suckling and suckling phase at their young age and would be more present eating solid feed from age d 30 and d 40 regardless the animal size. This could be explained by the fact that the solid feed was available at d 10 in the pen. The piglets were then neophobic to the new solid feed at d 15 and d 20 and then started to be used to it around d 30 and d 40. The frequency of suckling was higher when the piglets are young compared to when the piglets are older and looking for a source of feed elsewhere.

Post-weaning period

Soon after weaning, our first hypothesis was that the smaller, slower-growing piglets eat more roughage silage with high protein content available at the rack than the large piglets. Our results did not validate this first hypothesis and we showed that the Small piglets spent a major part of their time eating at the feeder filled with the phase 2 feed which was available during the lactation period, the Mediums were eating the wrapped alfalfa feed at the rack, and that the Large piglets stayed the major of their time out of the feeding zones. Moreover, the small piglets stayed longer in the feeding zone compared to the Large and Medium piglets. In that way, the present results did not confirm some preliminary study that reported that heterogeneity can be overcome by the ability of piglets to express compensatory growth in the presence of roughage silage with high protein content [START_REF] Bordes | Cahier Technique Alimentation des porcins en agriculture biologique[END_REF]. The sample size used in our study is relatively small (40 piglets and 5 litters) and could also explain the divergent findings between the studies. When considering the behavioural traits, only the total number of time passed in each zone was considered. It could have been interesting to look up also the number of visits or the average time spent exactly in each zone to have a comparison. The Large piglets were more present out of the zone of interest, this could also be explained that the behavioural video tests was done only the day after the weaning day and that the heavier piglets prefer resting or explore the straw in the comfortable kennel on straw bedding than eating. Plus, the videos observations at d 47 are available but not yet analyzed. Analyzing the videos could give us more information whether the presence of the small at the feeder, the medium at the rack and the heavier piglets out of the "feeding" zone are also confirmed at d47.

We confirmed the hypothesis that the small piglets would have a higher inflammatory state compared to the large piglets. Indeed, as expected, the small piglets had greater haptoglobin and oxidative production than Medium and Large piglets. The impacts of weaning on oxidative and inflammatory status have been already well documented [START_REF] Buchet | Effects of age and weaning conditions on blood indicators of oxidative status in pigs[END_REF], Sauwerwein et al., 2005, McCracken et al., 1995). It is not new that weaning had an impact on the physiology of the all piglets even though weaned at d 42 compared to d 21. We decided in the present study to check the BAP and the d-ROM indicator separately in order to have more information than combining them in a rate ratio [START_REF] Buchet | Effects of age and weaning conditions on blood indicators of oxidative status in pigs[END_REF]. As dominant-subordinate hierarchy is taking place at weaning period with new batches mixing unfamiliar piglets, the smaller piglets could have a higher inflammatory level by being the victims of social hierarchy by the presence of wound on the animal and by a greater stress level.

Our last hypothesis that the small piglets would have compensated their low body weight by catching up the heavier piglets was not validated. The average weight of the small, medium and Large piglets were significantly different from each other at d 42, this isn't surprising as we did in purpose to create weight group at d 42 to actually see compensation of weight at d 70. Yet, it is a little surprising to notice that the average weight of the small, medium and large piglets were also significantly different from each other at d 70. Indeed, regarding all the hypotheses concerning the feeding strategy, all were not valid. Moreover the smaller piglets were quite active during post weaning and could have used more energy compared to the heavier piglets which actually stayed the major of their time out of the feeding zone

CONCLUSION

Our results showed that the piglets with small body weight at weaning actually did not eat more solid feed during the lactating period and were not more interested by the roughage silage with high protein content during post-weaning. Additionally, they seemed to be in an inflammatory state as suggested by their greater plasma haptoglobin concentration. Altogether, these observations could explain that the Small piglets did not compensate their low body weight to catch up the larger piglets.

In the future, it could be interesting to focus on different points. It could be interesting to focus on the extreme piglets in term of body weight. Another perspective could be to look at those few piglets that actually did compensate their growth. For that purpose, a larger sample size would be necessary. Further analyses are already ongoing to study the metabolic status and the fecal microbiota which could also give more information on their adaptive strategies. Finally piglet adaptation weaned at 42 days in organic farming still need further research to understand the behavioural and physiological mechanisms involved in heterogeneous piglets' adaptation strategies.

Thanks also to my supervisor and to the scientists of INRA : Stéphane Ferchaud, Nathalie Le Floc'h, Marie-Christine Salaün and Lucile Montagne for sharing their expertise in the subject during all the duration of my thesis. They were always present and glad to help for improving my manuscript, finding solutions together to my encountered issues.

A special thanks to Stéphane Ferchaud who trusted me for this project and who lent me the project in total autonomy. He also taught me a lot in organic pig farming system. I also thank Rémi Resmond, the biostatiticien of INRA Pegase, for his patience and who explained me a lot about R script and statistical analysis.

Thanks to Carole Guerin for her assistance for Observer XT and thanks also to Lucien Yot, Lucas Fontaine (two interns) and the staff of experimental farm "Le Parquet" and "La Gouvanière" (Stéphane Moreau, Franck Guiraud, Tony Terasson, Yoann Bailly, Patrick Manceau and Doryan Grivault) for their help all along the experiments, as behavioural observations, animal care, laboratory work, camera installation, or arrangement of housing for the animals.

I acknowledge also my roommates (Marion Estrade, Nicolas Ferrand, Arthur Couturier, Fatemeh) with whom I had great time out of work and with whom we exchanged a lot about our different field of research at INRA. I acknowledge also my friend from France for their support: Amandine Piras, Julie Leblond, Alexis Carlier and Alice Scaillierez. I acknowledge Mathieu Carlier, for his support all along my thesis and my parents, my brothers and sisters for their precious support, their help improving my report and without whom I would not have got the opportunity to carry this Thesis for Wageningen University.

Figure 1 :

 1 Figure 1 : Photo of the camera recording during post weaning at d43 with the different zone recorded (Personal source) ......................................................................................................

Figure 2 :

 2 Figure 2: Box-plot representing weight of each category (Small, Medium and Large) at d42 ..................................................................................................................................................

Figure 3 :

 3 Figure 3: State-transition plot maps for category of piglets weight (Small, Medium, Large) between d 1 (Day 1 of age) and d 42 (Day 42 of age). Arrows show the direction of the transition with the population number which transited, absence of arrows means no transition ..................................................................................................................................................

Figure 4 :

 4 Figure 4 : Graph of the quantitative variables regarding the notation of the scan sampling done at d15, d20, d30 and d 40. (A: Pre-suckling and suckling phase; B= drink; C= Conspecific manipulation; E=Explore; I=Non-active; M= Eat; O= other including play ......

Figure 5 :

 5 Figure5: Individual factor map regarding the notation of the scan sampling done at d15 (J15), d20 (J20), d30 (J30) and d40(J40) in the morning (9h) and in the afternoon (3pm) .....

Figure 6 :

 6 Figure 6 : Representation of the Factor Analysis of Mixed Data (FAMD) of all piglets during the 8 scan sampling .................................................................................................................

Figure 7 :

 7 Figure 7 : Principal Component Analysis of all the variables regarding the video Analysis (with contrib = contribution to axis) ........................................................................................

Figure 9 :

 9 Figure 9 : Box plot of the Haptoglobin, BAP and d-Rom plasma concentrations for each weight category at d42 (Small, Medium and Large) at four different ages (d 42, d48, d 55, and d 62). .........................................................................................................................................

Figure 10 :

 10 Figure 10 : Evolution of the average weight of the 3 weight categories of piglet per week at post weaning .............................................................................................................................

  Ethogram of behaviors recorded on Piglets in the maternity unit adapted from Bowden and Staldler, 2008 ..................................................................................................... Table 2: Number of piglets from each category (Small, Medium and Large) at d42 present on the three different levels of teats positions (Anterior, Medium and Posterior) ........................

Figure 1 :

 1 Figure 1 : Photo of the camera recording during post weaning at d43 with the different zone recorded (Personal source)

Figure 2 :

 2 Figure 2: Box-plot representing weight of each category (Small, Medium and Large) at d42

Figure 3 :

 3 Figure 3: State-transition plot maps for category of piglets weight (Small, Medium, Large) between d 1 (Day 1 of age) and d 42 (Day 42 of age). Arrows show the direction of the transition with the population number which transited, absence of arrows means no transition

  represent the results of the Principal Component Analysis regarding the eight scan sampling observations done at d 15 (J15 in the figure 5), d 20 (J20), d 30 (J30) and d 40 (J40). Figure 4 represent the graph of the quantitative variables and Figure 5 represents the individual Factor map. The analysis of Figure 4 showed two clear principal axes explaining the variability of the dataset. Dimension 1 (Dim 1) represents inactive piglets on the left and active piglets on the right (explore, drink, conspecific manipulation, other behaviours including play behaviour). Dimension 2 (Dim 2) represents on top piglets which are more involved in the suckling phase and on the bottom those which prefer to eat the solid feed.

Figure 4 :Figure 5 :

 45 Figure 4 : Graph of the quantitative variables regarding the notation of the scan sampling done at d15, d20, d30 and d 40. (A: Pre-suckling and suckling phase; B= drink; C= Conspecific manipulation; E=Explore; I=Non-active; M= Eat; O= other including play

Figure 7 :Figure 8 :

 78 Figure 7 : Principal Component Analysis of all the variables regarding the video Analysis (with contrib = contribution to axis)

Figure 9 :

 9 Figure 9 : Box plot of the Haptoglobin, BAP and d-Rom plasma concentrations for each weight category at d42 (Small, Medium and Large) at four different ages (d 42, d48, d 55, and d 62).

Table 1 : Ethogram of behaviors recorded on Piglets in the maternity unit adapted from Bowden and Staldler, 2008

 1

ACKNOWLEDGEMENTS

I acknowledge my home school ISA Lille with Joop Lensink as supervisor and Wageningen University with Nicoline Soede as supervisor for giving me the opportunity to do my thesis in the Netherlands.

Thank you to the chair group Adaptation Physiology with Nicoline Soede who kindly accepted to be my supervisor from Wageningen, and who was always available and responsive by email, by taking news from the Netherlands and supervising my thesis. I also thank Kleijn, Lora vander, secretary of ADP Group who helped me with all the paperwork for the thesis and the colloquium.

Individual Growth Performance and health status

Growth performance

In order to ensure a precise follow-up of growth performance, the piglets were weighed individually on an automatic scale connected to the computer at d 1-10-21-28-42-48-55-62-70.

Biochemical and Immune Indicators

To characterise the physiological status of the piglets around weaning, blood samples (5mL Vacutainer EDTA and 5mL Heparin puncture tube at the jugular gulf) were taken from each piglet at weaning day (d 42) and then every week up to d 62 (d 48, d 55 and d 62). During the experiment, animals with clinical signs (diarrhea, weight loss, and poor general condition) were also collected, associated as far as possible with a representative healthy contemporary animal. The samples were then centrifuged at 2 500 g for 10 minutes at 4 ° C., aliquoted, and then stored at -20 ° C until the time of the biochemical analyzes at INRA Saint-Gilles. Biochemical analyzes were performed using the following measuring instruments: Konelab 20. Two oxidative statuses: Biological Antioxidant Potential (BAP) and Reactive Oxygen Metabolitesderived compounds (d-ROM) were measured as well as an inflammatory status (Haptoglobin).

Behavioural Observations

During the lactation period, all individual piglets (58 piglets) were observed for behavioural tests to characterise the "early status" of the piglets that will be observed after weaning. At d 10, the position of each piglet on the sow's teats was recorded as anterior teats, middle teats and posterior teats to check whether the small piglets were having milk from the posterior or the anterior teats of the sow.

Before the morning behavioural observation each litter was marked on their back with a form (horizontal line: -, or straight line: | or a dot: •) of different colours (black, blue, green, purple or red). During all the study, piglets always kept its own symbol. Eight scan sampling for behaviour observations were done at lactating period and two continuous behavioural video tests were done after weaning, but only one was analysed due to lack of time.

Result on the evolution of the piglets weight

Piglets were weighted at weaning day (d42) and then were categorized as Small, Medium and Large. Then piglets were weighted after the weaning at d48, d55, d62 and d70 of age.

The evolution of the average BW of the three weight categories of piglets after the weaning, illustrated in the Figure 10, was similar for the three categories with no compensatory growth for the Small piglets. The average BW between the three categories differs significantly at d 42 (p-value < 0.0001) andat d 70 (See Appendix 3 and4).

Appendix 1: Validation period of 4 hours instead of 12 hours for behavioural video observation

We notice that the activity level is similar for a period of observation of 4 hours and for 12 hours. We therefore considered the choice of observing the behavioural video for a period of 4 hours (between 2 and 6 pm) instead of 12h (between 8am and 8 pm) as valid.

Variability of piglet adaptability at weaning in organic farming system

In organic farming, weaning of piglets remains a critical period, although piglets are weaned older than in the conventional system. The use of hormones being forbidden and the lack of heat synchronization leads to spread of birth associated with heterogeneous ages, weight and health status of the piglets. The differences can be reinforced by a poorly adapted solid diet before and after weaning. The aim of the study was to understand the difference involved in the physiological mechanisms in piglet adaptation strategies to propose adapted feeding strategies. The studied population of fifty piglets born from five sows (Landrace x Duroc) of the INRA-Genesi unit and a boar (Piétrain x Large White). At weaning (d42), five batches of ten pigs per sow, were constituted by weight range: four "Large" piglets, two "Medium" and four "Small". They had access to a phase 2 feed from ten days and to roughage, high protein, after weaning (Wrapped alfalfa). Animals weights were recorded before and after weaning (d42-48-55-62). Blood samples were collected for inflammatory indicators and oxidative stress measurements (d42-48-55-62). Behavioural observations were video-recorded the day after weaning (d43) to establish weaning adaptation profiles. The small piglets spent on average more time in the area of feeding interest than the largest. The Small piglets also had a higher haptoglobin plasma concentration than the large and medium piglets, and there was no weight compensation for these piglets compared to the Medium and Large piglets. The average weight significantly differs between all three categories at d 42 and d70. Further analyses will permit to better characterize adaptative profiles of pig after the weaning.


domain: math info

text:
Object Identification

Object Identification

What is an identifier?

An identifier is a sort of name (could be alphanumeric or numeric only) that identifies a specific object (digital or not) in a set of objects. In an ideal world identifier should be unique for each object (bijection),

In practice this is rarely the case.

Object Identification

Non ambiguous

An identifier only stands for one resource. Whatever the database or source, two objects can not have the same identifier.

Confusions happens when different resources share the same identifier.

This characteristic is mandatory for identifiers.

How do we want to identify? good identifier.

ID36B2 ID36B2

Object Identification

Non ambiguous An identifier only stands for one resource. Whatever the database or source, two objects can not have the same identifier.

Confusions happens when different resources share the same identifier.

This characteristic is mandatory for identifiers.

How do we want to identify? good identifier.

Plot566 in 2016 Plot566 in 2017 can change over time: e.g. plot cutting

Object Identification

Non ambiguous Persistent

A persistent identifier is an identifier that is permanently assigned to an object (Ideally usable in several decades).

Aims : reusability of data over the long term (H2020 requirement)

The problem is that during periods of decades, many changes can occur within databases, but also in institutions or organizations in charge of the data.

It is thus necessary to preserve and recover dependencies between these elements, this in time and localisation.

How do we want to identify? good identifier.

Object Identification

Non ambiguous Persistent Resolvable (Dereferencable)

How do we want to identify? With an efficient identifier.

An identifier is said to be dereferenceable if it is possible to access the object or all the digital contents describing the object (e.g. URL, URI...).

Identification

What do we want to avoid for an efficient identification?

Non usability for operators:

  DOI or URI are string that identifies a particular resources DOI: 10.1111/nph.15385 → http://doi.org/10.1111/nph.15385 http://www.inra.fr/mp3/2015/arch/exp21/plant227

  traceability is the capability to trace data and each stage of their transformation. • keep track of the different versions Interoperability Interoperating in an efficient way: Working on quality level of interoperability Need to be maintained on the long term! take into account the persistence of human resources and skills Romain David, Laurence Mabile, Mohamed Yahia, Anne Cambon-Thomsen, Anne-Sophie Archambeau, et al.. How to assess FAIRness to improve crediting and rewarding processes for data sharing? A step forward towards an extensive assessment grid. RDA 13th (P13) Plenary Meeting, Apr 2019, Philadelphia, United States. ⟨https://www.rd-alliance.org/rda-13th-plenary-meeting-information⟩. ⟨10.5281/zenodo.2625721⟩. ⟨hal-02094678⟩

Interoperability Improvement locks: Too much standards and speed of standard evolution

		Interoperability	Copying identifiers without errors?
		Improvement needs:
	Interoperability ? Bad practices ?	Data format Data typology		• Architecture
		Shared semantics	• Data qualification
		Rate of database updates	• Taxonomic framework
				• Repository of actors
	Compatibility	Standard	Interoperability	• Conditions of use
	• Accessibility • Geographical repository • … • Too much semantic in the identifier (avoid optional metadata) Data format, Data typology, semantics & ontologies, Data qualification, Data standards… ○ (http://www.domainname.fr/m3p/Program/work_package/Country/Site/Year/Month/Day/Operator/P tools, technologies, groups...
	lant/Leave/Method/Color_of_the_pen/Length)

• No semantic in the identifier / excessively long ○ 10908538265365831680853826536583168 ○ http://www.domainname.fr/m3p/arch2017c17000915/FMfcgxwCgCSgPwdZxzkZtSHhnmvWWcpp • Confusing letters ○ Little L and maj i : I & l are not easily differentiable ○ (idem for O letter and 0 number) A & B are compatibles in another way than C & D : Harder now to interoperate A & B with C & D Interoperability is not only with your domain: Do not create your own format / ontology without prospecting communities approved standards (in & outside of your community) ?

Useful resources ARK:https://tools.ietf.org/html/draft-kunze-ark-18B2HANDLE: https://github.com/EUDAT-B2SAFE/B2HANDLECROP Ontology: https://github.com/bioversity/Crop-OntologyDOI: https://www.doi.org/ePIC: https://www.pidconsortium.eu/GUID: https://fr.wikipedia.org/wiki/Globally_Unique_IdentifierHandle System Namespace and Service Definition; http://www.ietf.org/rfc/rfc3651.txtHandle System Protocol (ver 2.1) Specification http://www.ietf.org/rfc/rfc3652.txtHTML: https://www.w3.org/html/ HTTP protocol: https://www.w3.org/Protocols/IRI: https://www.ietf.org/rfc/rfc3987.txt Linked data: https://www.w3.org/wiki/LinkedDataLSID: http://www.lsid.info/ ORCID: https://orcid.org/ OWL: https://www.w3.org/OWL/Plant Trait Ontology: https://github.com/Planteome/plant-trait-ontologyPURL: https://en.wikipedia.org/wiki/Persistent_uniform_resource_locatorRDF (W3C): https://www.w3.org/RDF/RDF-S: https://www.w3.org/TR/rdf-schema/SKOS: https://www.w3.org/TR/skos-reference/SPARQL: https://www.w3.org/TR/rdf-sparql-query/URI: https://www.w3.org/wiki/URIURL: https://www.w3.org/TR/url/URN: https://www.w3.org/urn/UUID: https://www.w3.org/wiki/UriSchemes/uuid XRI (OASIS): https://www.oasis-open.org/committees/xri/

Identification

Summary of what we want for an efficient identification

Agility needed for the long term maintenance!!

Data authority: a community approved and identified institution or body that is responsible of any type of action concerning data on the long term.

Variable naming

Naming rules

Plant Height

Naming rules

Plant Height

Interoperability

.

Interoperable Informations systems allow data exchange and reuse among scientific disciplines, organisations and countries through syntactically parseable and semantically understandable operations.

View publication stats View publication stats


Introduction: Whey protein aggregates for controlling emulsion heat-stability and texture

• Whey proteins :

-natural emulsifiers (adsorb to oil/water interface) -heat-sensitive proteins (heat treatments are required to extent food product shelf life)

→ heat sensitive emulsions → texturized (gelled) emulsions

Heat stability of whey protein emulsions (pH 7)

Stable/fluid How to design whey protein emulsions at high protein concentrations that are fluid after heating in the absence of non-dairy additives?

Results

Critical casein concentration

Observation at:

Oil

Results

How to obtain texturized (gelled) emulsions at low whey protein concentrations without non-dairy additives ? The number of « connectors » defines the texture of the emulsions (gel, viscous, fluid)

Main findings Introduction: whey proteins as texturizing ingredients

Whey proteins:

• Heat-induced aggregates produced by heating treatments in solution for increased functional properties

• Dry heating or heating a protein powder: process used for white egg powders for increasing foaming, How to custom their functional properties such as their ability to entrap large amounts of water and deliver high viscosities?

• Yield of transfer of whey proteins in microparticles with:  time of dry heating  addition of lactose (0.2, 0.3 and 0.5 g/10 g protein)

• 1 g of powder can lead to 0.9 g of microparticles General conclusions

Fluid•

  To design whey protein emulsions at high protein concentration (>4%) that are fluid after heating? • To design texturized (gelled) emulsions at low whey protein concentrations (<3%)?Research QuestionsHow to control the texture of whey protein emulsions in a large range of protein concentrations without non-dairy additives? whey protein emulsions at high protein concentrations that are fluid after heating in the absence of non-dairy additives?

-

  whey protein emulsions at high protein concentrations that are fluid after heating in the absence of non-dairy additives? -by adding sufficient amount of caseins to cover oil droplet surface (control of the size and stability of the oil droplets) → estimated value: mass of oil (g) × specific surface (m 2 /g of oil) × protein interfacial load (g/m 2 ) (protein interfacial load ∼ 2mg/m 2 for caseins) By selecting large and dense whey protein aggregates (small number, low interfacial adsorption rate and low reactivity on heating) STLOpen Days 19-21 March 2019 10 How to obtain texturized (gelled) emulsions at low whey protein concentrations without non-dairy additives ? Whey protein aggregates used as connectors (low density/elongated aggregates → factal aggregates) Size of the whey protein aggregates ≈ inter-droplet distance

  texturized (gelled) emulsions at low whey protein concentrations without non-dairy additives ? -Select aggregates of low density (fractal aggregates of whey proteins) -Use whey protein aggregates as « connector » at the surface of the oil droplets (size of the aggregates ∼ distance between oil droplets) -In combination with the homogenization pressure, use caseins as emulsifiers to control the size of the oil droplets (distance between oil droplets) « connected » oil droplets Whey protein aggregates (= connecting agent) Caseins (with the homogenizing pressure, they control the oil droplet size and the distance between oil droplets)

  emulsifying and gelling properties (Kato et al. 1989), • Dry heating of dairy powders: less studied and not used • Dry heating: a way to produce other changes of whey proteins  Limited diffusion of solutes = less aggregation  Traces or addition of lactose, degradation products of Maillard intermediates such as dicarbonyl  crosslinks

••

  • 1 g of powder leads to around 15 g of wet particles • The water content is around 95% of the wet weigh of microparticles • Due to the porous structure of powders Mechanism of formation and nature of the protein crosslinks is still speculative • We can form microparticles only at pH > 8.0 • The longer the whey protein solution is stored at pH 9.5, the higher the amount of microparticles • Role of:  alkaline pH  protein denaturation? Increased exposure of lysine residues?  lactose  Degradation via Maillard reactions? or degradation products of lactose at high temperature? Is still under study. Whey protein aggregates: high diversity of structures  High diversity of functionalities Large and dense aggregates (microgels) → heat-stability (solutions/emulsions)  Low density aggregates (fractal) → functionality dependent on the size o Small fractal aggregates : cold-gelation (CaCl 2 or acidic pH), limited syneresis o Medium fractal aggregates : connectivity between droplets, texture o Large fractal aggregates : increase viscosity  Very large porous aggregates (microparticles) → water entrapment, texture • Whey protein aggregates can be produced at the industrial scale (poster N°4 -Domitille de Guibert: "Designing innovative technological routes for the production and stabilization by drying of protein fractal assemblies").

How to custom protein assemblies with targeted sizes?

How to custom their functional properties such as their ability to entrap large amounts of water and deliver high viscosities?

• to replace food texturizing additives How to custom protein assemblies with targeted sizes?

How to custom their functional properties such as their ability to entrap large amounts of water and deliver high viscosities?

• Porous structure of powders  Microparticles entrap a large amount of water


Léa Salelles

Understanding the action mechanism of enzymes on solid substrate during food digestion

Keywords:

teaching and research institutions in France or abroad, or from public or private research centers.

Understanding the action mechanism of enzymes on solid substrate during food digestion

Lea Salelles

To cite this version: • Links between diet and health have been particularly highlighted these last years, especially in diseases appearing with aging (Parkinson, intestinal inflammatory diseases, sarcopenia, diabetes…).

• Growing market for « health oriented » food specifically designed for targeted nutritional needs.

What are the limiting factors in gastric digestion of protein gels by pepsin? What is the impact of the network microstructure on the kinetics of proteolysis in simulated gastric conditions?

• Innovative in vitro techniques to monitor the enzyme activity at microscopic and macroscopic scales on solid substrates.

• Microstructural investigations of the kinetics of gel proteolysis: imaging of local pH gradients, pepsin diffusion and gel breakdown during digestion of different protein substrates.

• A mathematical model of in vitro gastric digestion of protein gels.

• Transferring the developped methodologies to other heterogeneous catalysis systems.

• Predicting the digestion kinetics of more complex solid foods (with lipids and carbohydrates) by adapting the previous mathematical model.

• Designing protein-based food formulas adapted to specific populations.

Diffusion of acidic gastric fluid in an egg white gel, revealed with a pH indicator

Food structure appears as an interesting lever to control digestion, nutrient release and absorption. However, some mechanisms of action of digestive enzymes on solid food are still a subject of debate.

Complex protein matrices evolving during digestion

Acidification pH Enzyme Activity (pepsin)

In vivo Gastric conditions

Amino acids in blood

  Lea Salelles. Understanding the action mechanism of enzymes on solid substrate during food digestion. STLOpendays, Institut National de Recherche Agronomique (INRA). UMR UMR INRA / AgroCampus Rennes : Science et Technologie du Lait et de l'?uf (1253)., Mar 2019, Rennes, France. hal-02098689

  pH + Temperature + Time of reaction + Substrate (concentration and conformation, size of particles) + Water activity and ionic content Living Lab UR 0370 QuaPA