Counteract of bone marrow of blotchy mice against the increases of plasma copper levels induced by high-fat diets in LDLR−/− mice

BackgroundBone marrow of blotchy mouse (blotchy marrow) reflects the function of transmembrane domain and relevant intramembrane sites of ATP7A in myeloid cells. By chronic infusion of angiotensin II, we previously found that blotchy marrow plays a minor role in regulating plasma copper. Moreover, the recipients of blotchy marrow presented a moderate reduction of plasma lipids and inflammatory mediator production. Little is known about whether these changes are a specific response to angiotensin II or reveal a more general role of ATP7A.Objective and designWe investigated if blotchy marrow reduces plasma lipids and inflammatory mediators induced by high-fat diets. To test this hypothesis, blotchy and control marrows were reconstituted to the recipient mice (irradiated male LDLR−/− mice), followed by high-fat-diet feeding for 4 months. At the end points, plasma metals (copper, zinc and iron), lipid profiling (cholesterol, triglyceride, phospholipids and lipoprotein) and six inflammatory mediators (lymphotacin, MCP3, MCP5, TIMP1, VEGF-A and IP-10) were measured. Parallel experiments were performed using male LDLR−/− mice fed either high-fat diets or chow diets for 4 months.ResultsIn addition to hyperlipidemia and low-grade inflammation, high-fat diets selectively increased plasma copper concentration compared to chow diets in LDLR−/− mice. After high-fat-diet feeding, the recipients with blotchy marrow showed a decrease in plasma copper (p < 0.01) and an increase in plasma iron (p < 0.05). The recipients with blotchy marrow also presented decreases in cholesterol (p < 0.01) and phospholipids (p < 0.05) in plasma. Surprisingly, plasma levels of MCP3 (p < 0.05), MCP5 (p < 0.05), TIMP1 (p < 0.01), VEGF-A (p < 0.01) and IP-10 (p < 0.01) were significantly increased in the recipients with blotchy marrow compared to controls; the increased levels of MCP3, MCP5 and TIMP1 were more than 50%.ConclusionOur studies showed that blotchy marrow counteracts the increased copper levels induced by high-fat diets, indicating that circulating myeloid cells can regulate blood copper levels via ATP7A. Moreover, transplantation of blotchy marrow followed by high-fat diets leads to a decrease in lipid profile and an increase in inflammatory mediator production. Overall, blotchy marrow mediates divergent responses to angiotensin II and high-fat diets in vivo.     

The effect of age and gender on 38 chemical element contents in human iliac crest investigated by instrumental neutron activation analysis

ProjectTo understand the role of major, minor, and trace elements in the etiology of bone diseases including osteoporosis, it is necessary to determine the normal levels and age-related changes of bone chemical elements.ProcedureThe effect of age and gender on 38 chemical element contents in intact iliac crest of 84 apparently healthy 15–55 years old women (n=38) and men (n=46) was investigated by neutron activation analysis.ResultsMean values (M±SEM) for mass fraction (on dry weight basis) of Ca, Cl, Co, Fe, K, Mg, Mn, Na, P, Rb, Sr, and Zn for both female and male taken together were Ca – 169±3 g/kg, Cl – 1490±43 mg/kg, Co – 0.0073±0.0024 mg/kg, Fe – 177±24 mg/kg, K – 1820±79 mg/kg, Mg – 1840±48 mg/kg, Mn – 0.316±0.013 mg/kg, Na – 4970±87 mg/kg, P – 79.7±1.5 g/kg, Rb – 1.89±0.22 mg/kg, Sr – 312±15 mg/kg, and Zn – 65.9±3.4 mg/kg, respectively. The upper limit of mean contents of Cs, Eu, Hg, Sb, Sc, and Se were Cs≤0.09 mg/kg, Eu≤0.005 mg/kg, Hg≤0.005 mg/kg, Sb≤0.004 mg/kg, Sc≤0.001 mg/kg, and Se≤0.1 mg/kg, respectively. In all bone samples the contents of Ag, As, Au, Ba, Br, Cd, Ce, Cr, Gd, Hf, La, Lu, Nd, Sm, Ta, Tb, Th, U, Yb, and Zr were under detection limits.ConclusionsThe Ca, Mg, and P contents decrease with age, regardless of gender. Higher Ca, Mg, P, and Sr mass fractions as well as lower Fe content are typical of female iliac crest as compared to those in male bone.     

Dietary inclusion of raw faba bean instead of soybean meal and enzyme supplementation in laying hens: Effect on performance and egg quality

An experiment was conducted with 160 Hisex Brown laying hens to evaluate the effect of different inclusion levels of faba bean (FB) and enzyme supplementation on productive performance and egg quality parameters. The experimental diets consisted of five levels of FB: 0% (control), 25%, 50%, 75% and 100%, substituting soybean meal (SBM), and two levels of enzyme supplementation (0 or 250 mg/kg). Each dietary treatment was assigned to four replicate groups and the experiment lasted 22 weeks. A positive relationship (P < 0.05) was found between FB inclusion and body weight (BW) change of hens when compared to those of the control treatment. Enzyme supplementation significantly affected the final hens’ BW. Feed consumption (FC) of hens was statistically increased with increasing FB level up to 50%. Supplementing dietary enzyme mixture at 250 mg/kg led to improvement in FC at all studied ages (P < 0.05). Inclusion of 25% or 50% FB in diets had no adverse effects on feed conversion ratio (FCR) compared to the higher FB inclusion levels (75% or 100%). Egg weight (EW), egg number (EN) and egg mass (EM) were significantly (P < 0.05) influenced by FB inclusion in diet during the entire experimental periods, except for EN and EM at 20–24 weeks of age. Egg productive parameters were not influenced by enzyme mixture supplementation (P > 0.05). The main effect of FB levels replacing for SBM affected (P < 0.05) yolk and shell percentages, yolk index, yolk to albumen ratio, shell thickness and egg shape index. It can be concluded that FB and enzyme supplementation could be included in hens diet at less than 50% instead of SBM to support egg productive performance, however higher raw FB levels negatively affected egg production indices and quality.     

The effects of dietary boric acid and borax supplementation on lipid peroxidation,antioxidant activity,and DNA damage in rats

The aims of this study were to clarify the effects of high dietary supplementation with boric acid and borax, called boron (B) compounds, on lipid peroxidation (LPO), antioxidant activity, some vitamin levels, and DNA damage in rats. Thirty Sprague Dawley male rats were divided into three equal groups: the animals in the first group (control) were fed with a standard rodent diet containing 6.4 mg B/kg, and the animals in the experimental group were fed with a standard rodent diet added with a supra-nutritional amount of boric acid and borax (100 mg B/kg) throughout the experimental period of 28 days. The B compounds decreased malondialdehyde (MDA), DNA damage, the protein carbonyl content (PCO) level in blood, and glutathione (GSH) concentration in the liver, Cu–Zn superoxide dismutase (SOD), and catalase (CAT) activity in the kidney. The B compounds increased GSH concentration in blood and the vitamin C level in plasma. Consequently, our results demonstrate that B supplementation (100 mg/kg) in diet decreases LPO, and enhances the antioxidant defense mechanism and vitamin status. There are no differences in oxidant/antioxidant balance and biochemical parameters except for serum vitamin A and liver GSH concentration, between the boron compounds used in this study.     

Effect of copper sources and levels on serum lipid profiles in Black Bengal (Capra hircus) kids

Twenty eight 2–3 month old castrated male Black Bengal kids (Capra hircus) were used to determine the effects of dietary Cu concentration on lipid metabolism. These kids were randomly assigned to one of seven treatments in a ((2 × 3) + 1) factorial arrangement. Factors were two sources of Cu (CuSO₄ versus Cu proteinate) fed at three dietary levels (10, 20 or 30 mg/kg) and the control group, where neither CuSO₄ nor Cu proteinate were supplemented. Kids were fed a basal diet containing maize (19.5%), soybean (17.0%), deoiled rice bran (56.5%), molasses (4.0%), di-calcium phosphate and salt (1.0% each), and mineral and vitamin mixture (0.5% each) supplements, at 3.5% of BW to meet NRC requirements for protein, energy, macro minerals and micro minerals, excluding Cu. The basal diet (DM basis) contained 5.7 mg Cu/kg, 122.5 mg Fe/kg, 110 mg Zn/kg, 0.26 mg Mo/kg and 0.32% S. CuSO₄ or Cu proteinate (Cu-P) was added to the basal diet at the rate of 10, 20 and 30 mg/kg. Kids were housed in a well-ventilated shed with facilities for individual feeding in aluminum plated metabolic cages in an open-sided barn. Blood samples were collected on Days 0, 30, 60 and 90 to determine serum cholesterol, high density lipoprotein (HDL), total lipid and phospholipids. Kids were slaughtered after metabolism trial and liver tissues were collected to determine the copper and zinc concentrations. Kids receiving Cu-P showed higher (P < 0.05) HDL, total lipid and phospholipid concentrations. Increase in dietary level of Cu significantly decreased (P < 0.05) serum cholesterol and increased serum HDL, total lipid and phospholipid concentrations. There was an increasing (P < 0.05) trend in liver Cu with the increased dietary level of Cu supplementation irrespective of source, but the increasing rate was greater with CuSO₄ than Cu-P supplementation. Kids’ diet containing 30 mg/kg CuSO₄ had 26% more liver Cu than those fed iso-amounts of Cu-P. Fecal Cu excretion was increased with the increasing dietary level of Cu, and excretion was reduced by the use of Cu-P in the diet. In conclusion, dietary supplementation of organic Cu in the form of copper proteinate had significant effects on lipid metabolism in goat kids. There was an increase in accumulation of Cu in the liver and excretion of Cu in feces with the increase of dietary level of Cu in the diet of Black Bengal kids.     

Dietary preference for methionine sources in weaned pigs

The objective of the current study was to investigate the preference of weaned pigs given the choice of diets supplemented with dl-methionine (DLM) or liquid dl-methionine hydroxy analog-free acid (MHA-FA). A basal diet (BD) was formulated to contain 2.5 g methionine (Met) per kilogram of diet. The experimental diets included: (1) BD, (2) BD + 1 g DLM/kg, (3) BD + 2 g DLM/kg, (4) BD + 1.13 g MHA-FA/kg, (5) BD + 1.52 g MHA-FA/kg, (6) BD + 2.25 g MHA-FA/kg and (7) BD + 3.05 g MHA-FA/kg. Sixty weaned mixed-sex pigs were allotted to 5 treatment groups with 12 pig replicates per treatment in a randomized complete block design. During a 35-day experimental period, pigs in treatment group 1 received the BD whereas pigs in the other 4 treatment groups were allowed to choose between a pair of diets with either added 1 g DLM/kg in combination with either 1.13 or 1.52 g MHA-FA/kg or 2 g DLM/kg in combination with 2.25 or 3.05 g MHA-FA/kg, respectively. Pigs were housed in individual pens and had free access to feed and water. Daily feed intake (FI) was used as an indicator of diet preference. Cumulatively, pigs showed a preference (% of total FI) for the diet added with 1 g/kg DLM at 74% (P<0.05) in group 2, and 65% in group 3. Irrespective of the level of MHA-FA supplementation (2.25 or 3.05 g/kg), a preference for the diet supplemented with 2 g DLM/kg was 84% (P<0.05) in groups 4 and 5. During the entire period, pigs consistently (P<0.05) consumed more and preferred the diets supplemented with DLM more than the diets supplemented with MHA-FA in groups 2, 4 and 5. The preference for diets supplemented with DLM was more pronounced at higher Met supplementation levels. Feeding pigs a pair of diets supplemented with DLM or MHA-FA improved (P<0.05) the final body weight, daily weight gain, and FCR. The performance of pigs among the Met-supplemented groups was not different. In conclusion, when given a choice pigs preferred the diets supplemented with DLM more than the diets supplemented with liquid MHA-FA.     

Relative bioavailabilities of organic zinc sources with different chelation strengths for broilers fed diets with low or high phytate content

A six-day experiment was conducted to estimate the relative bioavailability values (RBV) of zinc (Zn) in three organic sources (oZn) with different chelation strengths compared to inorganic ZnSO₄ (iZn) for broilers fed a low or high phytate diet. A total of 1080, one-d-old male broiler chicks were randomly assigned to one of 18 dietary treatments (six replicates cages of ten chicks per cage) in a completely randomized design involving a 2 × 2 × 4 factorial arrangement with two levels of added phytate (0 or 10 g phytate as sodium phytate/kg), two levels of added Zn (30 or 60 mg/kg) and four Zn sources (iZn and three oZn sources) plus one low and one high phytate control treatments without Zn addition. The three oZn sources consisted of (1) Zn amino acid with weak chelation strength (ZnAA-L, formation quotient Q_f = 6.6, containing 119 g Zn/kg), (2) Zn proteinate with moderate chelation strength (ZnPRO-M, Q_f = 30.7, containing 133 g Zn/kg) or (3) Zn proteinate with strong chelation strength (ZnPRO-H, Q_f = 944.0, containing 186 g Zn/kg). Chicks were harvested at 6 days of age and pancreas metallothionein (MT) mRNA expression was used to estimate Zn RBV. Pancreas MT mRNA expression increased (P<0.01) as dietary Zn level increased. Chicks fed high phytate diets had lower (P<0.05) MT mRNA expression than chicks fed low phytate diets. Based on multiple linear regression slope ratios with ZnSO₄ set at 1.00, the RBV of ZnAA-L, ZnPRO-M and ZnPRO-H were 1.01, 1.28 and 0.70, respectively, for low phytate diets, and 1.05, 1.39 and 0.92, respectively, for high phytate diets. The slope for the oZn source with moderate chelation strength differed (P<0.05) from iZn and the other two oZn sources. The RBV of ZnAA-L, ZnPRO-M and ZnPRO-H under the high phytate diet increased by 0.04, 0.11 and 0.22, respectively, compared to those under the low phytate diet. Results indicate that the oZn sources with moderate or strong chelation strength offer partial or complete resistance to interference from high dietary phytate during digestion; and the oZn with moderate chelation strength had a greater RBV with both low and high phytate diets than iZn or oZn sources with weak or strong chelation strength.     

Promising role of ferulic acid,atorvastatin and their combination in ameliorating high fat diet-induced stress in mice

AimsThe present study evaluated a comparative and combined hepatoprotective effect of atorvastatin (AS) and ferulic acid (F) against high fat diet (HFD) induced oxidative stress in terms of hyperlipidemia, anti-oxidative status, lipid peroxidation and inflammation.Main methodsMale Swiss albino mice were given a diet containing high fat (H) (23.9% wt/wt), supplemented with AS (10 mg/kg) or F (100 mg/kg) and both (10 and 100 mg/kg) for 8 weeks. The control mice (C) were fed with normal diet.Key findingsThe H mice exhibited increased body weight; hyperlipidemia; serum level of tumor necrosis factor alpha (TNF-α), interleukin-6 (IL-6); hepatic lipid profile; lipid accumulation; reactive oxygen species (ROS) of hepatocytes, lipid peroxidation and liver antioxidant capacity was decreased. Immunofluorescent and Western blot assay revealed activation of nuclear factor kappa B (NF-κB) signaling pathway. The addition of F or AS and both in the diet significantly counteracted HFD induced body weight gain; hyperlipidemia; TNF-α, IL-6; hepatic lipid profile; fatty infiltration; NF-κB signaling pathway; ROS; lipid peroxidation and moreover elevated levels of hepatic antioxidant enzymes activity were observed.SignificanceSimultaneous treatment with AS, F and their combination protected against HFD induced weight gain and oxidative stress. The protection may be attributed to the hypolipidemic and free radical scavenging activity of AS or F and their combination. This study illustrates that AS and F have relatively similar hypolipidemic, antioxidative, anti-inflammatory actions and the AS + F combination along with HFD has shown outstanding effects as compared to other treated groups.     

Influence of supplemental enzymes on the performance and phosphorus excretion of broilers fed wheat-based diets to 6 weeks of age

Efficacies of phosphorolytic enzymes (phytase+acid phosphatase), and an enzymic “cocktail” (phytase+acid phosphatase+pectinase+citric acid), were investigated in broilers fed wheat-based diets from day 1 to 43. Broilers were fed the following four diets: (1) a positive control diet (7.1 g total P/kg, 4.1 g non-phytate P/kg, 9.8 g Ca/kg); (2) a low phosphorus diet (4.1 g total P/kg, 1.7 g non-phytate P/kg, 6.0 g Ca/kg) supplemented with phytase (750 units/kg) and acid phosphatase (3156 units/kg); (3), a low phosphorus diet (4.1 g total P/kg, 1.7 g non-phytate P/kg, 6.0 g Ca/kg) supplemented with phytase, acid phosphatase, pectinase (1900 units/g) and citric acid (20 g/kg); and (4) a low phosphorus diet (4.1 g total P/kg, 1.7 g non-phytate P/kg, 8.0 g Ca/kg) supplemented as in diet 3. For the grower period (22–43 days), the contents of P and Ca were lowered by 0.2 and 0.3 g/kg, respectively. The dietary treatments were fed to three floor pen replicates of 50 birds each. For the starter period, there were no differences observed among dietary treatments in terms of body weight gains or feed efficiency. Total body weight gains for the starter and grower periods did not differ among dietary treatments, but total feed efficiency was significantly enhanced in birds fed diet 2. At the completion of the experiment chickens fed phosphorolytic enzymes had the best feed efficiencies, the highest contents of ash in the toes, and the highest carcass yield. Chicken receiving the cocktail of enzymes and 8 g Ca/kg (diet 4) performed as well as birds in the control treatment, but had higher yields of carcass and excreted 56% less phosphorus.     

Effects of supplementation with vitamin E on the performance and the tissue peroxidation of broiler chicks and the stability of thigh meat against oxidative deterioration

Two experiments were conducted: Expt 1 determined the optimal allowance of vitamin E in the diet for broiler chicks aged 0–3 weeks; Expt 2 investigated the effects of different dietary levels of vitamin E (α-tocopherol) on the performance and the oxidative stability of thigh meat of broiler chicks during storage. In Expt 1, 1-day-old 900 broiler chicks were allocated to five treatments, each with six replicates (cages) of 22 as-hatched chicks for performance evaluation, and another cage of 45 male chicks for determining plasma and hepatic α-tocopherol and thiobarbituric acid reactive substances (TBARS) concentration in blood and liver. The basal dietary α-tocopherol concentration was 13 mg/kg, and the five α-tocopherol acetate supplementation levels were 0, 5, 10, 50 and 100 mg/kg. For 0–3-week-old broiler chicks fed with maize–soya bean meal–soya oil type diet, supplementation of vitamin E did not influence the feed intake, but tended to improve growth and feed utilization, however there was no significant correlation between performance and vitamin E supplementation level. Significant positive correlations existed between dietary supplemental vitamin E level and plasma or hepatic α-tocopherol concentrations (P<0.05), and a negative correlation with hepatic TBARS levels no matter at what age (11, 16 and 21 days). In Expt 2, 2200 broiler chicks were randomly allocated to five treatments with four replicates (pens) in each. Chicks were fed ad libitum five pellet diets supplemented with vitamin E at 5, 10, 20, 50 and 100 mg/kg of diet, respectively. The basal dietary α-tocopherol level of grower and finisher diets were 7 and 6 mg/kg, respectively. Supplementation of vitamin E tended to improve growth and feed utilization of birds during 0–3 weeks of age, but the performance from 0 to 6 weeks of age were not influenced. The hepatic α-tocopherol concentrations of 6-week-old chicks linearly increased with the dietary vitamin E levels (R²=0.98, P<0.001). The content of TBARS in the thigh meat over 4 days of storage under 4°C was significantly decreased by increasing dietary vitamin E level (P<0.05). There was a significant inverse relationship between TBARS value in the thigh meat and the dietary vitamin E level (R²=0.93, P<0.01). Supplementation of vitamin E significantly improved the meat quality stability substantially against oxidative deterioration. Comparing the hepatic α-tocopherol levels of chicks in Expts 1 and 2, total allowance of dietary α-tocopherol of 20–30 mg/kg could sustain relatively constant hepatic α-tocopherol level at round about 2–2.5 μg/kg.     

Baicalin upregulates the genetic expression of antioxidant enzymes in Type-2 diabetic Goto-Kakizaki rats

AimThe primary purpose of this study was to characterize and investigate the antioxidant and anti-diabetic activities of the flavonoid baicalin in type 2 diabetic Goto-Kakizaki rats.Main methodsFour groups of Goto-Kakizaki rats (n = 6) were subjected to the following oral treatments for 30 days: (1) metformin — 500 mg/kg (2) baicalin — 120 mg/kg (3) metformin 500 mg/kg and baicalin — 120 mg/kg (4) vehicle treated diabetic controls receiving distilled water. The plasma glucose, triglyceride, total cholesterol, lipid peroxide and protein carbonyl contents were measured on a weekly basis. Following the completion of the treatment, the rats were sacrificed and their blood, heart, pancreatic and hepatic tissues were collected for analysis. The antioxidant enzyme activities as well as their expression were quantified using Western Blot, microarray and RT-PCR.Key findingsThe respective analyses showed that the baicalin- and the metformin and baicalin-treated groups had statistically significant increases (p < 0.05) in the activity and expression of the antioxidant enzymes (superoxide dismutase, catalase and glutathione peroxidase) compared with vehicle- and metformin-treated groups. Further complementing the antioxidant enzyme activity increases, the oxidative stress markers of plasma lipid peroxide and protein carbonyl contents were reduced in these groups as well. These treatment groups also had reduced plasma total cholesterol and triglyceride levels compared with vehicle-treated and metformin-treated groups (p < 0.05).SignificanceBaicalin was an efficient antioxidant in reducing hyperglycemia-induced oxidative stress through the increased expression of antioxidant enzyme activities. It was also an efficient anti-hypertriglyceridemic as well as anti-hypercholesterolemic agent compared with metformin.     

Effects of maternal treatment of dehydroepiandrosterone (DHEA) on serum lipid profile and hepatic lipid metabolism-related gene expression in embryonic chickens

Over the last decade, much evidence emerged to suggest that alterations in maternal diets during pregnancy may irreversibly affect aspects of physiological and biochemical functions in the fetus. To explore the effects of maternal dietary treatments with dehydroepiandrosterone (DHEA) on lipid metabolism in the embryo, we investigated serum lipid profile and hepatic lipid metabolism-related gene expression in the maternal and embryonic chicken. Sixteen-week-old pullets were allocated into 3 groups (n = 30), and after laying, they were provided with a commercial diet supplemented with DHEA at 0, 20 or 100 mg/kg diet. Eggs were collected after DHEA treatment and incubated at 37.5 °C and a relative humidity of 60%. Blood and liver samples were collected from hens and embryonic chickens. DHEA treatment resulted in decreased body weight and increased relative liver weight in both maternal and embryonic chickens, while the concentrations of blood triglyceride (TG), total cholesterol (TC), low-density lipoprotein-cholesterol (LDL-C) and non-esterified fatty acid (NEFA) were significantly lower in the 20 mg DHEA/kg group as compared to the control group during embryonic development. The expression of acetyl CoA carboxylase (ACC) and carnitine palmitoyl transferase I (CPTI) gene was also reduced following treatment with 20 mg DHEA/kg at hatching. However, blood TC, and hepatic fatty acid synthase (FAS) and hydroxy methylglutaryl-CoA reductase (HMGR) gene expression were significantly up-regulated in the 100 mg DHEA/kg group during embryonic development and hatching. Overall, the results of this study indicate that maternal dietary treatment with DHEA regulates serum lipid metabolism and hepatic gene expression.     

The effect of feeding expeller-pressed canola meal on growth performance and diet nutrient digestibility in weaned pigs

The effects of feeding increasing levels of expeller-pressed (EP) canola meal in substitution for soybean meal as an energy and amino acid source were evaluated in 240 weaned pigs with an initial body weight of 7.3 ± 0.6 kg. Five pelleted wheat-based diets containing 0, 50, 100, 150 or 200 g EP canola meal/kg were formulated to contain 10.0 MJ net energy (NE)/kg and 1.18 g standardised ileal digestible (SID) lysine/MJ NE and were fed for 4 wk starting 1 wk after weaning at 19 days of age. Expeller-pressed canola meal was added at the expense of soybean meal and the diets were balanced for NE using canola oil and for amino acids using crystalline lysine, methionine, threonine and tryptophan. Increasing inclusion of EP canola meal linearly reduced (P<0.001) the apparent total tract digestibility of energy, dry matter and crude protein and the digestible energy content of diets. From 0 to 28 days on trial, increasing inclusion of EP canola meal did not affect body weight gain, feed intake and feed efficiency. In conclusion, up to 200 g EP canola meal/kg can replace soybean meal in diets formulated to equal NE and SID amino acid content and fed to nursery pigs starting 1 wk after weaning without reducing growth performance.     

Application of a method to determine ileal digestibility in broilers of amino acids in wheat

Growing male chickens (Ross) were fed diets of increasing wheat content (250, 500 and 750 g/kg) in which wheat was the only protein source. Other dietary ingredients were: oil (50 g/kg), a vitamin and mineral premix (50 g/kg) and a 50 : 50 mixture of pure maize starch and glucose to make diets up to 1000 g/kg. Titanium dioxide was included at 5 g/kg as an inert marker. Four wheat samples were evaluated: varieties B and H each with a low and high crude protein content (102 and 130, respectively ; 96 and 128 g CP/kg at 880 g DM/kg. Experimental diets were fed ad libitum for 3 days from 18 days of age to birds housed in pairs in cages in a fully randomised design comprising 6 replicates per diet. Samples of ileal digesta were obtained from the birds following slaughter at 21 days of age and analysed for titanium and amino acid content. For each amino acid, the amount of apparently digestible amino acid in the diet was regressed against the dietary wheat content and the linear regression thus derived extrapolated to 1000 and 0 g wheat/kg, allowing the estimation of the apparently digestible amino acid content of the wheat sample and the endogenous loss of amino acids, respectively. Dividing the former by the total amino acid content of the wheat allowed calculation of the coefficient of apparent amino acid digestibility (CAD). Addition of the intercept to the amount of apparently digestible amino acid in the wheat sample and dividing this by the total amount of amino acid in the wheat allowed the determination of the coefficient of true digestibility (CTD). Values for methionine for the four wheat samples ranged between 0.749 to 0.842 (CAD) and 0.719 to 0.892 (CTD), and for lysine from 0.531 to 0.697 (CAD) and 0.510 to 0.793 (CTD). Results revealed that an increase in crude protein in the wheat varieties resulted in a significant increase in apparently digestible amino acid content (P < 0.001), for example, the digestible cystine contents of wheat were 1.54 and 2.22 g/kg for wheat H at the low and high crude protein contents, respectively. Coefficients of digestibility also appeared to increase as the crude protein increased.     

Hepatoprotective and antioxidant activity of N-Trisaccharide in different experimental rats

ObjectivesTo investigate the hepatoprotective, antioxidant and antihyperlipidemic effect of N-Trisaccharide isolated from Cucumis prophetarum (L.) on different experimental rats.MethodsN-Trisaccharide (25 and 50 mg/kg.b.w), silymarin (25 mg/kg) and glibenclamide (25 mg/kg) was orally administered once daily for 28 days and toxicity evaluation studies were carried out. Liver damage was assessed by determining DNA damage, serum enzyme activities and hepatic histopathology of carbon tetrachloride (CCl₄) induced hepatic injury in rats. Enzymatic and non enzymatic antioxidant levels in liver and kidney were determined and biochemical parameters such as, serum lipid profile, renal function markers were estimated in type 2 diabetic rats.ResultsDNA fragmentation analysis revealed the protective effect of N-Trisaccharide on liver DNA damage. Histopathological studies indicated that CCl₄-induced liver injury was less severe in N-Trisaccharide (25 and 50 mg/kg) treated group. Given at the above doses conferred significant protection against the hepatotoxic actions of CCl₄ in rats, reducing serum markers like SGOT, SGPT, ALP, creatinine and urea levels back to near normal (p < 0.05) compared to untreated rats. In diabetic rats, N-Trisaccharide treatment significantly reversed abnormal status of enzymatic and non-enzymatic antioxidants levels to near normal. Also, serum lipids such as TG, TC, LDL-C and VLDL-C levels were significantly (p < 0.05) reduced compared to diabetic untreated rats.ConclusionPresent study results confirm that N-Trisaccharide possesses significant antihyperlipidemic, antioxidant and hepatoprotective properties.     

Dietary supplementation of green synthesized manganese-oxide nanoparticles and its effect on growth performance,muscle composition and digestive enzyme activities of the giant freshwater prawn Macrobrachium rosenbergii

The green synthesized Mn₃O₄ nanoparticles (manganese-oxide nanoparticles) using Ananas comosus (L.) peel extract was characterized by various techniques. HR-SEM photograph showed that manganese-oxide nanoparticles (Mn-oxide NPs) were spherical in shape, with an average size of 40–50 nm. The Zeta potential revealed the surface charge of Mn-oxide NPs to be negative. Further, the Mn-oxide NPs were dietary supplemented for freshwater prawn Macrobrachium rosenbergii. The experimental basal diets were supplemented with Mn-oxide NPs at the rates of 0 (control), 3.0, 6.0, 9.0, 12, 15 and 18 mg/kg dry feed weight. The as-supplemented Mn-oxide NPs were fed in M. rosenbergii for a period of 90 days. The experimental study demonstrated that prawns fed with diet supplemented with 3–18 mg Mn-oxide NPs/kg shows enhanced (P < 0.05) growth performance, including final weight and weight gain (WG). Significant differences (P < 0.05) in feed conversion ratio (FCR) were observed in prawn fed with different diets. Additionally, prawns fed with 3.0–18 mg/kg Mn-oxide NPs supplemented diets achieved significant (P < 0.05) improvement in growth performance, digestive enzyme activities and muscle biochemical compositions, while, the prawns fed with 16 mg/kg of Mn-oxide NPs showed enhanced performance. Prawns fed on diet supplemented with 16 mg/kg Mn-oxide NPs showed significantly (P < 0.05) higher total protein level. The antioxidants enzymatic activity (SOD and CAT) metabolic enzymes status in muscle and hepatopancreas showed no significant (P > 0.05) alterations in prawns fed with 3.0–18 mg/kg of Mn-oxide NPs supplemented diets. Consequently, the present work proposed that 16 mg/kg of Mn-oxide NPs could be supplemented for flexible enhanced survival, growth and production of M. rosenbergii. Therefore, the data of the present study recommend the addition of 16 mg/kg of Mn-oxide NPs diet to developed prawn growth and antioxidant defense system.     

Pretreatment with a combination of quercetin and α-tocopherol ameliorates adenosine triphosphatases and lysosomal enzymes in myocardial infarcted rats

AimsMembrane bound adenosine triphosphatases (ATPases) and lysosomal enzymes play an important role in the pathology of myocardial infarction. This study was aimed to evaluate the combined preventive effects of quercetin and α-tocopherol on membrane bound ATPases and lysosomal enzymes in isoproterenol induced myocardial infarcted rats.Main methodsMale Wistar rats were pretreated with a combination of quercetin (10 mg/kg) and α-tocopherol (10 mg/kg) daily for 14 days. After the pretreatment period, isoproterenol (100 mg/kg) was injected to rats at an interval of 24 h for two days to induce myocardial infarction. The activities of ATPases and lysosomal enzymes were assayed.Key findingsIsoproterenol treated rats showed decreased levels of heart creatine kinase and lactate dehydrogenase. The activity of sodium potassium adenosine triphosphatase was decreased and the activities of magnesium adenosine triphosphatase and calcium adenosine triphosphatase were increased in isoproterenol treated rats. Also, the activities of β-glucuronidase, β-N-acetylglucosaminidase, β-galactosidase, cathepsin-B and D were increased (serum and heart), but the activities of β-glucuronidase and cathepsin-D were decreased in lysosomal fraction and increased in cytosolic fraction of the heart in isoproterenol treated rats. Furthermore, the heart lipid peroxidation products were increased in isoproterenol treated rats. Combined pretreatment with quercetin and α-tocopherol to isoproterenol treated rats normalized all the biochemical parameters studied. The observed effects are due to their membrane stabilizing property and this property might be due to decreased lipid peroxidation.SignificanceOur study demonstrated that combined pretreatment was better than single pretreatment. This study may have significant impact on myocardial infarcted patients.     

Effect of selenium-enriched malt on VEGF and several relevant angiogenic cytokines in diethylnitrosamine-induced hepatocarcinoma rats

One hundred and ninety-three Sprague–Dawley (SD) rats (average body weight 100–120 g) were randomly divided into five groups (I–V). Groups I and II rats served as the negative and positive controls respectively and both received 0.1 mg/kg Se from sodium selenite supplemented diets for the 18-week experimental period. Groups III–V rats were fed Se from SEM supplemented diets (0.3, 1 and 3 mg/kg respectively). To induce hepatocarcinoma, groups II–V rats received diethylnitrosamine solution (100 mg/L) at the dosage of 10 mg/kg body weight in drinking water daily for 16 weeks, followed by sterilized water for a further 2 weeks. Group I rats received sterilized water throughout. At weeks 4, 8, 12 and 16 five rats in each group were sacrificed by cervical decapitation. At the termination of the study, at week 18, the surplus rats were sacrificed by cervical decapitation. Feed was withheld from the rats for 12  h before sampling. The following items including TNF-α, IGF-II, NO and T-NOS levels in plasma were tested using kit techniques. At the same time the expression of vascular endothelial growth factor (VEGF) in tumor tissue was analyzed by immunohistochemistry using the envision two-step methods with a kit. The results indicated that SEM could increase the levels of TNF-α in the early stages of hepatocarcinoma formation, however there was a decrease in the later stage of hepatocarcinogenesis. SEM could also significantly decrease the levels of IGF-II and NO, and inhibit the expression of VEGF in tumor tissue. SEM delayed the development of hepatocarcinoma in rats and that could be partially attributed to inhibition of angiogenesis.     

Effects of exogenous enzymes on apparent nutrient digestibility in rainbow trout (Oncorhynchus mykiss) fed diets with high inclusion of plant-based protein

Plant-based protein ingredients are increasingly used in feed for carnivorous fish as replacement for fish meal. This study investigated if supplementing diets with high inclusion levels of plant-based protein with three different enzymes could improve the apparent nutrient digestibility in juvenile rainbow trout (Oncorhynchus mykiss). Three diets with high inclusion levels of either de-hulled, solvent extracted soybean meal (344 g/kg), sunflower meal (246 g/kg), or rapeseed meal (264 g/kg) were produced and feed batches were coated post-extrusion with the different enzymes: β-glucanase at 67 mg/kg, xylanase at 208 mg/kg, protease at 228 mg/kg, or a combination of the three enzymes at the same doses. Three consecutive digestibility trials were carried out using a flow-through, modified Guelph System. Each trial was designed to include five dietary treatment groups: a non-supplemented control diet and 4 diets supplemented with either β-glucanase, xylanase, protease or a combination of the three enzymes. Diets were fed to triplicate tanks and two faecal sampling periods for digestibility measurements for each tank were scheduled. Each experiment comprised a 10-day digestibility trial succeeded by a water sampling period for measuring the dissolved nitrogen (N) output. Each experiment lasted 17–19 days in total.Apparent digestibility coefficients (ADCs) of protein, fat, ash, phosphorus and dry matter (DM) were derived from the three digestibility trials, along with calculations of the specific growth rate (SGR, %/d) and feed conversion ratio (FCR). Nitrogen mass-balance and energy retention were evaluated for each dietary treatment group to elucidate on the utilization of digested nutrients and energy.Enzyme supplementation had only moderate effect on apparent nutrient digestibility in the sunflower and rapeseed experiments, while β-glucanase and protease improved the apparent digestibility of all dietary nutrients in the soybean experiment (P<0.05). The effect was more pronounced for lipid than for other nutrients. β-Glucanase had a positive effect on energy retention in the soybean experiment (P<0.05), while there were no effects on nitrogen retention or fish performance in any of the three experiments (P>0.05) during the short feeding periods. The study thus provides preliminary results on the potential of β-glucanase and protease to increase apparent nutrient digestibility of soybean meal in fish feed.     

High-selenium lentil diet protects against arsenic-induced atherosclerosis in a mouse model

Background: Cardiovascular disease (CVD) is a major cause of death worldwide, and arsenic (As) intake, mainly through drinking water, is a well-known risk factor for CVD as well as other health problems. Selenium (Se) is a known antagonist to As toxicity. Objective: We tested the potential of high-Se lentils from the Canadian prairies as a therapeutic food to alter the outcome of As-enhanced atherosclerosis. Materials and Methods: Male ApoE^−/− mice exposed to a moderate level of As (200 ppb) in their drinking water, and control mice on tap water received one of three lentil diets: Se-deficient (0.009 mg/kg), Se-adequate (0.16 mg/kg) or Se-high (0.3 mg/kg). After 13 weeks, lesion formation in the aortic arch and sinus were assessed. Intralesional cellular composition, serum lipid levels and hepatic oxidative stress were assessed as well. Results: Arsenic-exacerbated plaque formation was reduced in the sinus and completely abolished in the aortic arch of mice on the Se-fortified lentil diet, whereas lesions were increased in As-exposed mice on both the Se-deficient and Se-adequate diets. Notably, Se deficiency contributed to proatherogenic composition of serum lipids in As-exposed mice as indicated by high-density lipoprotein:low-density lipoprotein. At least adequate Se status was crucial for counteracting As-induced oxidative stress. Conclusion: This study is the first to show the potential of high-Se lentils to protect against As-triggered atherosclerosis, and this invites further investigations in human populations at risk from As contamination of their drinking water.