The potential of herbal extracts containing bioactive compounds to strengthen immunity could contribute to reducing antimicrobial use in poultry. This study aimed at developing a reliable and robust methodological pipeline to assess the ability of herbal extracts to strengthen chicken innate defenses, especially concerning inflammation and oxidative stress. This methodology was applied to Melissa officinalis L. (MEL) extract, recognized for its biological activities including antioxidant and anti-inflammatory properties. Different methods were used to (1). guarantee the quality of MEL extract and its capacity to stimulate the innate immune system; (2). evaluate the relevance of an ex vivo model to mimic inflammatory and oxidative stress challenges to replace LPS injection in chickens; (3). analyse the effects of feed supplemented with MEL extract on inflammation and oxidative stress induced ex vivo; (4). assess the effects of MEL extract on the redox balance, health, welfare and performance in broilers exposed to suboptimal starting conditions through a large-scale approach. The quality of MEL extract preparations, through phytochemical quantification of rosmarinic acid (RA), revealed varying concentrations of RA in the different MEL extracts. RA concentrations remained stable for at least 9 months and in feed three months after incorporating MEL extract. When incubated with chicken cell lines MEL extract showed potential metabolic activation and ability to stimulate immune functions but induced cytotoxicity at high concentrations. The original ex vivo model of inflammation developed on chicken blood cells enabled inflammation and oxidative stress biomarkers to be expressed and revealed antioxidative and anti-inflammatory properties of blood cells from chickens fed MEL extract. The experimental model of chicken suboptimal starting conditions validated beneficial effects of MEL extract on the redox balance and also evidenced improved performance during the growth phase, a tendency for fewer muscle defects but a higher severity of pododermatitis lesions without affecting other welfare indicators. This study grouped methods and tools that could be combined according to the plant extract, the needs of professionals working in poultry production systems and staff responsible for animal health, welfare and feeding.
Induced by overfeeding, hepatic steatosis is a process exploited for the “foie gras” production in mule ducks. To better understand the mechanisms underlying its development, the physiological responses of mule ducks overfed with corn for a duration of 11 days were analyzed. A kinetic analysis of glucose and lipid metabolism and cell protection mechanisms was performed on 96 male mule ducks during overfeeding with three sampling times (after the 4th, the 12th, and the 22nd meal). Gene expression and protein analysis realized on the liver, muscle, and abdominal fat showed an activation of a cholesterol biosynthetic pathway during the complete overfeeding period mainly in livers with significant correlations between its weight and its cholesterolemia ( r = 0.88; P < 0.0001) and between the liver weight and the hmgcr and soat1 expression ( r = 0.4, P < 0.0001 and r = 0.67; P < 0.0001, respectively). Results also revealed an activation of insulin and amino acid cells signaling a pathway suggesting that ducks boost insulin sensitivity to raise glucose uptake and use via glycolysis and lipogenesis. Cellular stress analysis revealed an upregulation of key autophagy-related gene expression atg8 and sqstm1( P < 0.0001) during the complete overfeeding period, mainly in the liver, in contrast to an induction of cyp2e1( P < 0.0001), suggesting that autophagy could be suppressed during steatosis development. This study has highlighted different mechanisms enabling mule ducks to efficiently handle the starch overload by keeping its liver in a nonpathological state. Moreover, it has revealed potential biomarker candidates of hepatic steatosis as plasma cholesterol for the liver weight.
Kinetic study of the expression of genes related to hepatic steatosis, global intermediate 1 metabolism and cellular stress during overfeeding in mule ducks 2 3 ABSTRACT 24 25 Induced by overfeeding, hepatic steatosis is a reversible process exploited for "foie gras" 26 production. To better understand the mechanisms underlying this non-pathological 27 phenomenon, we analysed the physiological responses of the mule duck to cope with 22 28 carbohydrate meals. A kinetic analysis of intermediate metabolism and cell protection 29 mechanisms was performed during overfeeding. As expected, dietary carbohydrates are up 30 taken mainly by the liver (chrebp, glut1/2/8) and converted into lipids (acox, scd1, acsl1, fas, 31 dgat2). Our study showed an activation of cholesterol biosynthetic pathway with significant 32 correlations between plasma cholesterol, expression of key genes (hmgcr, soat1) and liver 33 weight. Results revealed an activation of insulin and amino acid cell signalling pathway 34 suggesting that ducks boost insulin sensitivity to raise glucose uptake and use via glycolysis 35 and lipogenesis. Expression of cpt1a, acad, hadh suggested an induction of beta-oxidation 36 probably to remove part of newly synthesized lipids and avoid lipotoxicity. Cellular stress 37 analysis revealed an upregulation of autophagy-related gene expression (atg8, atg9, sqstm1) in 38 contrast with an induction of cyp2e1 suggesting that autophagy could be suppressed. Lamp2a 39 and plin2 enhanced, conflicting with the idea of an inhibition of lipophagy. Hsbp1 40 overexpression indicated that mechanisms are carried out during overfeeding to limit cellular 41 stress and apoptosis to prevent the switch to pathological state. Atf4 and asns overexpression 42 reflects the nutritional imbalance during overfeeding. These results permitted to highlight the 43 mechanisms enabling mule ducks to efficiently handle the huge starch overload and reveal 44 potential biomarker candidates of hepatic steatosis as plasma cholesterol for liver weight. 45
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