The Unique Features of Proteins Depicting the Chicken Amniotic Fluid.

In many amniotes, the amniotic fluid is depicted as a dynamic milieu that participates in the protection of the embryo (cushioning, hydration, and immunity). However, in birds, the protein profile of the amniotic fluid remains unexplored, even though its proteomic signature is predicted to differ compared with that of humans. In fact, unlike humans, chicken amniotic fluid does not collect excretory products and its protein composition strikingly changes at mid-development because of the massive inflow of egg white proteins, which are thereafter swallowed by the embryo to support its growth. Using GeLC-MS/MS and shotgun strategies, we identified 91 nonredundant proteins delineating the chicken amniotic fluid proteome at day 11 of development, before egg white transfer. These proteins were essentially associated with the metabolism of nutrients, immune response and developmental processes. Forty-eight proteins were common to both chicken and human amniotic fluids, including serum albumin, apolipoprotein A1 and alpha-fetoprotein. We further investigated the effective role of chicken amniotic fluid in innate defense and revealed that it exhibits significant antibacterial activity at day 11 of development. This antibacterial potential is drastically enhanced after egg white transfer, presumably due to lysozyme, avian beta-defensin 11, vitelline membrane outer layer protein 1, and beta-microseminoprotein-like as the most likely antibacterial candidates. Interestingly, several proteins recovered in the chicken amniotic fluid prior and after egg white transfer are uniquely found in birds (ovalbumin and related proteins X and Y, avian beta-defensin 11) or oviparous species (vitellogenins 1 and 2, riboflavin-binding protein). This study provides an integrative overview of the chicken amniotic fluid proteome and opens stimulating perspectives in deciphering the role of avian egg-specific proteins in embryonic development, including innate immunity. These proteins may constitute valuable biomarkers for poultry production to detect hazardous situations (stress, infection, etc.), that may negatively affect the development of the chicken embryo.

Egg serpins: The chicken and/or the egg dilemma.

Twenty-seven serpins belonging to clade A, B, C, D, E, F, G, H and I serpins are currently referenced in chicken genome databases. Phylogenetic analysis of chicken serpins revealed that ovalbumin (Serpinb14) and its paralogs ovalbumin-related protein Y (Serpinb14b) and ovalbumin-related protein X (Serpinb14c) are found in bird species. These clade B serpins are specifically expressed in reproductive tissues and exported in the egg where they constitute major protein components. These data suggest that these three paralogs have probably appeared in birds to face new environments and ensure the extra-uterine development of an embryo in a shell egg. Twelve other serpins have been identified in the newly produced egg, some of them having a specific distribution in the respective egg structures (eggshell, egg white, vitelline membrane and egg yolk). The physiological role of these egg serpins remain largely unexplored, but there is increasing evidence in literature or by homologies with their mammalian counterparts, that some of them participate in cell proliferation, tissue remodeling and/or angiogenesis associated with folliculogenesis and development of extraembryonic structures, eggshell biomineralization, egg defense and nutrition of the embryo. A better knowledge of the phylogenetic evolution of these 15 serpins in other oviparous species, on their egg distribution, on their regulation during embryonic development (activation/degradation/transfer) and on their functional specificity, is needed to better appreciate their role and their bird-specificity. These review shed light on the multiple possibilities that offer the avian egg model to study the role of serpins in reproduction and developmental biology.

The use of active PET to package rosé wine: Changes of aromatic profile by chemical evolution and by transfers.

Active Polyethylene Terephthalate (PET) bottles containing 1 or 3% of oxygen scavenger (named 1osPET and 3osPET) were used to pack rosé wine. Changes in the aromatic profiles were monitored during 12months and compared to those of a wine packed in glass bottles. Wine in 1osPET bottles was differentiated from wine in glass or 3osPET bottles by ten aging markers such as cis-dioxane, ethyl pyruvate or furfural. Only trans-1,3-dioxolane allowed to discriminate wine in glass and in 3osPET bottles. Methionol, an oxygen sensitive aroma compound, was preserved in glass and 3osPET bottles but was slightly degraded (15%) in 1osPET bottles. Chemical reactions were the main cause of the aroma compound degradation. Indeed, the total amount of compounds sorbed only reached 160µg considering the bottles and the joint of cap after 12months of storage. The use of PET with 3% of oxygen scavenger is adapted to pack wine for at least 12months.

Aromatic evolution of wine packed in virgin and recycled PET bottles.

The evolution of the aromatic profile of a rosé wine packed in glass, virgin and recycled PET bottles was studied. Wine stored in PET and glass bottles was clearly differentiated after 5months of storage but only by a limited number of compounds. More pronounced decrease of oxygen sensitive compounds such as methionol was observed in PET bottles as well as the apparition of oxidative and ageing aroma compounds such as ethyl pyruvate, furfural or dioxanes in higher concentration. Compared to virgin PET bottles, recycled PET bottles induced slight changes favouring the presence of esters and alcohols. The chemical evolution of wine was the most important phenomenon that explains the loss of flavour rather than the sorption into PET. Because of their moderate oxygen permeability, the use of virgin PET and recycled PET bottles could be adapted for short conservation of wine but detrimental to aromatic quality if long conservation is intended.

Effect of ethanol on the sorption of four targeted wine volatile compounds in a polyethylene film.

The objectives of this study were to demonstrate that the presence of ethanol in a solution containing two esters and two aromatic alcohols has several consequences on the sorption of these compounds into polyethylene (PE) film. First, sorption of ethanol into the PE film occurred at the same time as water and reached 8 kg m(-3) using 12% v/v of ethanol. This sorption was associated with an increase in PE crystallinity, which may have prevented the sorption of volatile compounds despite their strong affinity with PE film, as evaluated by Hansen solubility parameters. Moreover, increasing the ethanol concentration increased the solubility of the four volatile compounds. In the case of aromatic alcohols, the sorption was decreased in the presence of ethanol as expected. In the case of esters, as their hydrolysis was substantial in the presence of water, the consequence was a higher sorption into the PE film in the presence of ethanol than in its absence. Nevertheless, the sorption also depended on the concentration of ethanol and the heterogeneity of the ethanol-water mixture as well as the presence of other volatile compounds, as in the case of 4-ethylphenol. In conditions simulating wine packaging, losses of volatile compound by sorption and by permeation estimated after only 5 days of contact varied between 0.08 and 25% for 2-phenylethanol and ethyl hexanoate, respectively.