Identification of Differentially Expressed Proteins in Liver in Response to Subacute Ruminal Acidosis (SARA) Induced by High-concentrate Diet.

The aim of this study was to evaluate protein expression patterns of liver in response to subacute ruminal acidosis (SARA) induced by high-concentrate diet. Sixteen healthy mid-lactating goats were randomly divided into 2 groups and fed either a high-forage (HF) diet or a high-concentrate (HC) diet. The HC diet was expected to induce SARA. After ensuring the occurrence of SARA, liver samples were collected. Proteome analysis with differential in gel electrophoresis technology revealed that, 15 proteins were significantly modulated in liver in a comparison between HF and HC-fed goats. These proteins were found mainly associated with metabolism and energy transfer after identified by matrix-assisted laser desorption ionization/time of flight. The results indicated that glucose, lipid and protein catabolism could be enhanced when SARA occurred. It prompted that glucose, lipid and amine acid in the liver mainly participated in oxidation and energy supply when SARA occurred, which possibly consumed more precursors involved in milk protein and milk fat synthesis. These results suggest new candidate proteins that may contribute to a better understanding of the mechanisms that mediate liver adaptation to SARA.

Changes in milk performance and hepatic metabolism in mid-lactating dairy goats after being fed a high concentrate diet for 10 weeks.

Feeding a high concentrate (HC) diet is a widely used strategy for supporting high milk yields, yet it may cause certain metabolic disorders. This study aimed to investigate the changes in milk production and hepatic metabolism in goats fed different proportions of concentrate in the diet for 10 weeks. In total, 12 mid-lactating goats were randomly assigned to an HC diet (65% concentrate of dry matter, n=6) or a low concentrate (LC) diet (35% concentrate of dry matter, n=6). Compared with LC, HC goats produced greater amounts of volatile fatty acids and produced more milk and milk lactose, fat and protein (P<0.01). HC goats showed a greater concentration of ATP, NAD, plasma non-esterified fatty acids and hepatic triglycerides than LC goats (P<0.05). Real-time PCR results showed that messenger RNA (mRNA) expression of gluconeogenic genes, namely, glucose-6-phosphatase, pyruvate carboxylase and phosphoenolpyruvate carboxykinase were significantly up-regulated and accompanied greater gluconeogenic enzyme activities in the liver of HC goats. Moreover, the expression of hepatic lipogenic genes including sterol regulatory element-binding protein 1c, fatty acid synthase and diacylglycerol acyltransferase mRNA was also up-regulated by the HC diet (P<0.05). HC goats had greater hepatic phosphorylation of AMP-activated protein kinase than LC (P<0.05). Furthermore, histone-3-lysine-27-acetylation contributed to this elevation of gluconeogenic gene expression. These results indicate that lactating goats fed an HC diet for 10 weeks produced more milk, which was associated with up-regulated gene expression and enzyme activities involved in hepatic gluconeogenesis and lipogenesis.

High concentrate-induced subacute ruminal acidosis (SARA) increases plasma acute phase proteins (APPs) and cortisol in goats.

The aim of this study was to investigate changes of stress status in dairy goats induced to subacute ruminal acidosis (SARA). The level of acute phase proteins (APPs) including haptoglobin (HP) and serum amyloid A (SAA) in plasma and their mRNA expression in liver, as well as plasma cortisol and genes expression of key factors controlling cortisol synthesis in adrenal cortex were compared between SARA and control goats. SARA was induced by feeding high concentrate diet (60% concentrate of dry matter) for 3 weeks (SARA, n=6), while control goats (Con, n=6) received a low concentrate diet (40% concentrate of dry matter) during the experimental time. SARA goats showed ruminal pH below 5.8 for more than 3 h per day, which was significantly lower than control goats (pH>6.0). SARA goats demonstrated a significant increase of hepatic HP and SAA mRNA expression (P<0.05), and the level of HP but not SAA in plasma was markedly increased compared with control (P<0.05). The level of cortisol in plasma showed a trend to increase in SARA goats (0.050.05). These results suggested that SARA goats experienced a certain stress status, exhibiting an increase in HP production and cortisol secretion.

The effect of equol injection in ovo on lipid metabolism and hepatic lipogenic gene expression in broilers.

This study investigated the effects of in ovo administration of equol (Eq) on post-hatch growth and hepatic lipid metabolism in broiler chickens. Fertilized eggs (146 eggs/group) were injected with 0 µg (control, Con), 20 µg (low dose, L) and 100 µg (high dose, H) Eq in the albumen on the 7th day of incubation. Except a trend increase in the weight of total fat (P = 0.09), Eq had no effect on growth or liver weight in broilers at 49 days of age. Males presented higher liver and BWs and lower total fat and relative liver weights than females (P < 0.01). However, there were no significant effects of Eq or Eq-gender interactions on growth performance or tissues weight (P > 0.05). With respect to lipid parameters in the serum, the results showed that female broilers presented higher triacyglycerol (TG) and low-density lipoprotein cholesterol concentrations than males, whereas there was no gender difference in serum total cholesterol (TC) or high-density lipoprotein cholesterol (HDLC) concentration (P > 0.05). Eq administration significantly decreased serum TG and TC but increased HDLC concentrations in serum of broilers at 49 days of age (P < 0.05), whereas there were no interactions between gender and Eq (P > 0.05). To elucidate the mechanism behind the significant changes of serum TG and TC levels, the expression of genes involved in lipid metabolism in the liver was investigated in female chickens using reverse transcription-PCR. Carnitine palmitoyl transferase I (CPTI) messenger RNA (mRNA) was significantly upregulated by 20 and 100 µg Eq (P < 0.05). High-dose Eq significantly decreased fatty acid synthase (FAS) and enhanced cholesterol-7alpha-hydroxylase (CYP7A1) mRNA levels in the liver (P < 0.05). Eq had no significant effects on acetyl-CoA carboxylase, sterol regulatory element binding protein-1c, malic enzyme, low-density lipoprotein receptor or 3-hydroxy-3-methylglutaryl coenzyme A reductase mRNA in the liver (P > 0.05). These results in female broilers suggest that Eq decreased blood TG by upregulating CPTI and downregulating FAS mRNA expression in the liver, and that high serum cholesterol levels stimulated CYP7A1 gene transcription in the liver.

The effect of equol injection in ovo on posthatch growth, meat quality and antioxidation in broilers.

In order to investigate the long-term effects of equol (Eq) on growth and meat quality in broilers, 0 µg (control, Con), 20 µg (low dose, L) and 100 µg (high dose, H) Eq, respectively, were injected into fertile eggs (146 eggs per group) on 7 days of embryos. After hatch, chickens were fed under the same conditions and slaughtered at 49 days of age for sample collection and analysis. The results showed that body weight and composition were marginally affected by Eq administration (P > 0.05). Compared with their male counterparts, the meat quality of female broilers was affected greatly after Eq administration. The redness (a*) of meat color in the L and H groups of female broilers was significantly decreased by 24.10% and 21.50% (P < 0.01), respectively; cooking loss decreased by 12.11% and 16.82%, respectively, in the L and H groups (P < 0.01); 24 h and 48 h drip loss was significantly decreased by 60.27% and 45.72% (P < 0.05), respectively, in the H group. However, for male broilers, only cooking loss was significantly decreased by high dosage of Eq treatment (P < 0.05). The antioxidative status was analyzed for discovering further the mechanism behind the improvement of the water-holding capacity caused by Eq in female broilers. The activity of glutathione peroxidase (GSHPx) in plasma was greatly increased by 15.94% in the L group (P < 0.01), whereas the total superoxide dismutase activity (T-SOD) and the content of malondialdehyde in plasma were not changed (P > 0.05). The T-SOD activity in the breast muscle of the L and H groups were significantly improved by 23.14% and 18.82% (P < 0.05), respectively. GSHPx in the breast muscle of the H group showed a tendency to increase (P = 0.06 < 0.1). These results indicate that Eq injection in ovo does not affect the growth of broilers, but significantly improves the water-holding capacity of the muscle, especially in female broilers, which is related to the improvement of antioxidative status.

Effect of daidzein on egg-laying performance in Shaoxing duck breeders during different stages of the egg production cycle.

The effect of a phyto-oestrogen, daidzein, on the laying performance of Shaoxing female ducks was examined in three experiments performed at different stages of the egg production cycle. Egg-laying rate, egg weight, egg composition, feed conversion ratio, hatchability characteristics of eggs and body weight, ovary and oviduct weight, as well as changes in serum concentrations of T3, T4 and E2 were recorded as response criteria. In the first experiment, 68 young ducks, 100 d of age, were given a basal diet (maize-soybean meal) with or without 3 mg of daidzein/kg diet for 42 d. Daidzein did not affect the onset of lay but apparently decreased egg-laying rate and mean egg weight as well as the feed conversion ratio. In the second experiment, 240 breeding ducks, 402 d of age, were allotted at random to three groups and given the basal diet containing daidzein at 0 (control), 3 (Da1) and 5mg/kg (Da2) for 35d. Egg-laying rate, mean egg weight and feed conversion ratio increased in both Da1 and Da2 groups. However, an adverse effect of daidzein on fertility and hatchability was observed. In the third experiment, 320 breeding ducks, 415 d of age, were fed on the basal diet with or without 5mg of daidzein/kg diet for 63 d. Egg-laying rate increased by 7.7%, average egg weight tended to increase, whereas yolk/albumen ratio decreased. Daidzein-treated ducks had higher body weight and oviduct weight compared with their controls. Elevated plasma T4 and E2 concentrations accompanied these phenotypic changes, but serum T3 was not affected. It is suggested that daidzein exerts divergent effects on the egg-laying performance of Shaoxing ducks under different physiological conditions and this action is dose-dependent. The changes in circulating E2 imply possible participation of endogenous oestrogen in the mechanism of daidzein action.