Electrochemical Determination of Levodopa Using Zinc Sulfide Nanospheres-Reduced Graphene Oxide.

Zinc sulfide nanospheres (ZnS NSs) were prepared by hydrothermal synthesis and graphene oxide (GO) was prepared by the Hummer's method. ZnS NSs-rGO/ITO electrode was synthesized by heat treatment at a certain temperature, which was used for the detailed electrochemical determination of levodopa (LD). Finally, they were annealed to form the ZnS NSs-rGO/ITO electrode for detecting levodopa (LD). The results reveal that the ZnS NSs with the diameter of ~1 µm are covered by rGO. The ZnS NSs-rGO/ITO electrode has a good sensitivity of 1.43 µA µM -1 for the determination of LD in the concentration range of 1-40 µM. Moreover, it also shows a good selectivity, reproducibility and stability. In order to verify the practicability, we also use the electrode to detect LD in human serum. The detection results also prove that the electrode can be used in real life.

Metabolites of intestinal microflora upregulate miR-192-5p to suppress proliferation of colon cancer cells via RhoA-ROCK-LIMK2 pathway.

OBJECTIVE: The application of intestinal microflora is involved in various cancers; however, researches reporting the potential of metabolites of intestinal microflora (MIM) on biological activities of colon cancer (CC) cells are unavailable. This study was designed to testify the functions of MIM on CC cells and its mechanism. MATERIALS AND METHODS: qRT-PCR/Western blot were applied to test the expression levels of miR-192-5p and BMPR2 in human colonic epithelial cells and CC cells (HCT116, SW480). The effects of MIM, mimics-miR-192-5p or inhibitors-miR-192-5p on mRNA and protein expressions of miR-192-5p and BMPR2 were verified by qRT-PCR and Western blot. MTT assay for CC cell viability, flow cytometry for CC cells apoptosis rate, and cell scratch and cell chamber served for the analysis of invasion and migration ability of CC cells. The relationship between miR-192-5p and BMPR2 was validated employing Luciferase reporter gene assay. RESULTS: Compared with human normal colonic epithelial cells, HCT116 and SW480 cells had lower expression of miR-192-5p and higher expression of BMPR2 (p < 0.01). MIM and mimics-miR-192-5p could enhance cell apoptosis and suppress the migration and proliferation of CC cells. MIM were also found to up-regulate miR-192-5p and down-regulate the expression levels of BMPR2 and p-LIMK2 (p < 0.01). Transfection of inhibitors-miR-192-5p reversed the inhibitory effect of MIM on CC cells. CONCLUSIONS: MIM could up-regulate miR-192-5p to inhibit CC cell growth via down-regulating BMPR2 and inhibiting the activity of RhoA-ROCK-LIMK2 pathway.

Effects of dietary gallic acid supplementation on performance, antioxidant status, and jejunum intestinal morphology in broiler chicks.

Gallic acid (GA), widely distributed in plants and feeds, is known to have a diverse range of activities such as anti-oxidant, anti-inflammatory, antibacterial, anti-allergic, anti-mutagenic, and anti-carcinogenic. The purpose of this study was to investigate the efficacy of inclusion of dietary GA at levels 0, 25, 50, 75, 100, or 150 mg/kg on growth performance, antioxidant status, and jejunum intestinal morphology of broiler chicks. In total, 630 one-day-old Arbor Acres (AA) male broiler chicks were randomly allotted to 6 treatment groups for a period of 6 weeks. The results indicate that dietary addition of GA at 75 to 100 mg/kg improved feed conversion efficiency in both the grower (d 21 to 42, P = 0.045) and overall (d 1 to 42, P = 0.026) periods. Dietary addition of GA at a concentration ≥100 mg/kg was able to exhibit higher breast muscle ratio at 42 d (P = 0.043). Interestingly, dietary GA inclusion level from 50 to 100 mg/kg reduced the crypt depth (P = 0.009) and increased the villus height:crypt depth ratio (VCR) of the birds (P = 0.006). Dietary supplementation of GA at 100 mg/kg decreased plasma malondialdehyde (MDA) content at 42 d of age (P = 0.030). Moreover, dietary addition of GA linearly increased plasma total antioxidant capacity (P = 0.039) and plasma total superoxide dismutase activities (P = 0.049) at 21 d of age. However, analysis of plasma biochemical markers revealed that dietary supplementation of GA did not exhibit beneficial health effects. Overall, we conclude that 75 to 100 mg/kg of GA are suitable for enhanced growth performance and health benefits in a broiler diet.

Effect of dietary supplementation of organic or inorganic zinc on carbonic anhydrase activity in eggshell formation and quality of aged laying hens.

This study evaluated the effects of different dietary levels and sources of zinc (Zn) on performance and carbonic anhydrase (CA) activity in eggshell formation and quality in aged laying hens. A total of 504 Hy-line Grey layers aged 59 wk were fed a basal diet (Zn, 28.4 mg/kg) for 4 wks, then randomly allocated to 7 groups that were fed a basal diet or a basal diet supplemented with inorganic (ZnSO4·H2O) or organic (amino acid metals, 9.58%) Zn at 35, 70, or 140 mg Zn per kg of feed for 6 weeks. Each group had 6 replicates of 12 hens. Results showed that egg weight decreased linearly with the supplemental level of organic Zn (P < 0.05). Dietary Zn supplementation had linear and quadratic effects on the CA activity in plasma (P < 0.05), and it was higher in the organic Zn-added groups at wks 2 and 4 (P < 0.05). Dietary Zn supplementation had a quadratic effect on the CA activity in the eggshell gland (P < 0.05). Shell thickness was greater in the organic Zn-added groups (P < 0.05), and its relationship with the supplemental level of Zn showed linearly and quadratically, increasing with the organic Zn and with the inorganic Zn at wk 4, while linearly increasing with the inorganic Zn at wk 6 (P < 0.05). At wk 4, the supplemental level of inorganic Zn had a linear effect on shell weight, and linear and quadratic effects on shell index and ratio (P < 0.05), while shell weight, the index, and ratio increased linearly and quadratically with the organic Zn level in the diet (P < 0.05), with more obvious effects in the organic Zn-added groups (P < 0.05). Overall, dietary Zn supplementation, up to 140 mg/kg feed, could increase eggshell thickness by enhancing CA activity in the plasma and eggshell gland of aged layers; thicker eggshells were found in the organic Zn-added groups, but the breaking strength did not increase despite the eggshell thickness increasing.

Effect of different protein ingredients on performance, egg quality, organ health, and jejunum morphology of laying hens.

The aim of this study was to evaluate the effect of different protein ingredients on performance, egg quality, organ health, and jejunum morphology of laying hens. A total of 216 32-wk-old Hy-Line W36 laying hens with similar egg production was randomly divided into 3 treatment groups with 6 replicates of 12 birds each. The experimental diets were isocaloric (metabolizable energy, 2 655 kcal/kg) and isonitrogenous (crude protein, 16.5%). The control group was fed a corn-soybean meal basal diet (SBM), and the other 2 experimental diets consisted of a basal diet with 195.0 g/kg low-gossypol cottonseed meal (LCSM) or 292.0 g/kg double-zero rapeseed meal (DRM). The feeding trial lasted 12 weeks. The egg production, daily feed intake, egg weight, and feed conversion ratio were not affected (P > 0.05) by diets used. The daily egg mass was decreased in the LCSM group in wk 7 to 12 (P < 0.05). Albumen height, Haugh unit, albumen weight, albumen proportion, and oviduct index were reduced in the 195.0 g/kg LCSM at wk 12 (P < 0.05), while yolk color was increased (P < 0.05). A lower yolk weight, yolk proportion, abdominal fat, dry matter of the liver, and fat content of the liver were observed in the DRM group at wk 12 (P < 0.05). Using LCSM or DRM as a sole protein ingredient in the diet reduced the ovomucin content of albumen when stored 28 d at 4°C. The villous height of the jejunum in the LCSM group was the lowest among treatments (P < 0.05). The mucosal columnar epithelial cell layer of the magnum in the LCSM group was incomplete, and there was a little albumen secretions in the lumen of the magnum. Together, our results suggest that the 195.0 g/kg LCSM diet may produce an adverse effect on daily egg mass and albumen quality. The adverse effect might be associated with the reduction of absorption ability in the jejunum and secretion capacity in the magnum.

Effects of dietary grape proanthocyanidins on the growth performance, jejunum morphology and plasma biochemical indices of broiler chicks.

Grape proanthocyanidins (GPCs) are a family of naturally derived polyphenols that have aroused interest in the poultry industry due to their versatile role in animal health. This study was conducted to investigate the potential benefits and appropriate dosages of GPCs on growth performance, jejunum morphology, plasma antioxidant capacity and the biochemical indices of broiler chicks. A total of 280 newly hatched male Cobb 500 broiler chicks were randomly allocated into four treatments of seven replicates each, and were fed a wheat-soybean meal-type diet with or without (control group), 7.5, 15 or 30 mg/kg of GPCs. Results show that dietary GPCs decrease the feed conversion ratio and average daily gain from day 21 to day 42, increase breast muscle yield by day 42 and improve jejunum morphology between day 21 and day 42. Chicks fed 7.5 and 15 mg/kg of GPCs show increased breast muscle yield and exhibit improved jejunum morphologies than birds in the control group. Dietary GPCs fed at a level of 15 mg/kg markedly increased total superoxide dismutase (T-SOD) activity between day 21 and day 42, whereas a supplement of GPCs at 7.5 mg/kg significantly increased T-SOD activity and decreased lipid peroxidation malondialdehyde content by day 42. A supplement of 30 mg/kg of GPCs has no effect on antioxidant status but adversely affects the blood biochemical indices, as evidenced by increased creatinine content, increased alkaline phosphatase by day 21 and increased alanine aminotransferase by day 42 in plasma. GPC levels caused quadratic effect on growth, jejunum morphology and plasma antioxidant capacity. The predicted optimal GPC levels for best plasma antioxidant capacity at 42 days was 13 to 15 mg/kg, for best feed efficiency during grower phase was 16 mg/kg, for best jejunum morphology at 42 days was 17 mg/kg. In conclusion, GPCs (fed at a level of 13 to 17 mg/kg) have the potential to be a promising feed additive for broiler chicks.

Dietary supplementation of pyrroloquinoline quinone disodium protects against oxidative stress and liver damage in laying hens fed an oxidized sunflower oil-added diet.

The protective effects of dietary pyrroloquinoline quinone disodium (PQQ.Na2) supplementation against oxidized sunflower oil-induced oxidative stress and liver injury in laying hens were examined. Three hundred and sixty 53-week-old Hy-Line Gray laying hens were randomly allocated into one of the five dietary treatments. The treatments included: (1) a diet containing 2% fresh sunflower oil; (2) a diet containing 2% thermally oxidized sunflower oil; (3) an oxidized sunflower oil diet with 100 mg/kg of added vitamin E; (4) an oxidized sunflower oil diet with 0.08 mg/kg of PQQ.Na2; and (5) an oxidized sunflower oil diet with 0.12 mg/kg of PQQ.Na2. Birds fed the oxidized sunflower oil diet showed a lower feed intake compared to birds fed the fresh oil diet or oxidized oil diet supplemented with vitamin E (P=0.009). Exposure to oxidized sunflower oil increased plasma malondialdehyde (P<0.001), hepatic reactive oxygen species (P<0.05) and carbonyl group levels (P<0.001), but decreased plasma glutathione levels (P=0.006) in laying hens. These unfavorable changes induced by the oxidized sunflower oil diet were modulated by dietary vitamin E or PQQ.Na2 supplementation to levels comparable to the fresh oil group. Dietary supplementation with PQQ.Na2 or vitamin E increased the activities of total superoxide dismutase and glutathione peroxidase in plasma and the liver, when compared with the oxidized sunflower oil group (P<0.05). PQQ.Na2 or vitamin E diminished the oxidized sunflower oil diet induced elevation of liver weight (P=0.026), liver to BW ratio (P=0.001) and plasma activities of alanine aminotransferase (P=0.001) and aspartate aminotransferase (P<0.001) and maintained these indices at the similar levels to the fresh oil diet. Furthermore, oxidized sunflower oil increased hepatic DNA tail length (P<0.05) and tail moment (P<0.05) compared with the fresh oil group. Dietary supplementation of PQQ.Na2 or vitamin E decreased the oxidized oil diet induced DNA tail length and tail moment to the basal levels in fresh oil diet. These results indicate that PQQ.Na2 is a potential antioxidant and is as effective against oxidized oil-related liver injury in laying hens as vitamin E. The protective effects of PQQ.Na2 against liver damage induced by oxidized oil may be partially due to its role in the scavenging of free radicals, inhibiting of lipid peroxidation and enhancing of antioxidant defense systems.

Dietary Protein Sources Affect Internal Quality of Raw and Cooked Shell Eggs under Refrigerated Conditions.

This study was conducted to evaluate the effects of various protein sources (soybean meal, SBM; cottonseed protein, CSP; double-zero rapeseed meal, DRM) on the internal quality of refrigerated eggs. A total of 360 laying hens (32 wk of age) were randomly allotted to six treatment groups (five replicates per treatment) and fed diets containing SBM, CSP, or DRM individually or in combination with equal crude protein content (SBM-CSP, SBM-DRM, and CSP-DRM) as the protein ingredient(s). A 6×3 factorial arrangement was employed with dietary types and storage time (0 d, 2 wk, and 4 wk) as the main effects. After 12 wk of diet feeding, a total of 270 eggs were collected for egg quality determination. The egg Haugh unit (HU) in the CSP, SBM-DRM, and DRM groups were significantly lower than those in the SBM and SBM-CSP groups. The hardness and springiness of the cooked yolk in the CSP group were significantly higher than those in the other treatment groups. A lower HU, lower yolk index and higher albumen pH were observed in the DRM group compared to the SBM and SBM-CSP groups when the eggs were stored to 4 wk, and the HU was improved in the CSP-DRM group compared to the DRM group (p<0.05). Higher yolk hardness was observed in the CSP group compared to the other groups during storage (p<0.05), but the hardness of the cooked yolk in the SBM-CSP and CSP-DRM groups showed no difference in comparison to the SBM group. In conclusion, CSP may ameliorate the negative effects of DRM on the HU of refrigerated eggs, and SBM or DRM may alleviate the adverse effects of CSP on yolk hardness.

Application of low-gossypol cottonseed meal in laying hens' diet.

An experiment was conducted to evaluate the application of dietary low-gossypol cottonseed meal (LCSM) in layers' diets. A total of 432 40-week-old Hy-line W36 laying hens were allocated to one of the six dietary treatments with 6 replicates of 12 birds each. The control group was fed a corn-soybean meal basal diet, and the 4 experimental diets consisted of a basal diet with 50, 98.3, 144.2, or 189 g/kg LCSM, respectively (correspondingly 25%, 50%, 75%, 100% dietary protein provided by soybean meal were replaced by LCSM). The sixth group was fed a basal diet supplemented with free gossypol (FG group). The feeding trial lasted for 12 weeks. The results showed that no significant difference was observed on egg production or feed conversion ratio (FCR, feed/egg, g/g) among all groups, but feed intake and egg weight were significantly decreased in the 189 g/kg LCSM group in weeks 46 to 51 (P < 0.05). A significant decrease was not observed in shell strength, shell thickness, and yolk color in all periods, but Haugh unit, albumen height, and egg white protein content were reduced in 189 g/kg LCSM group in weeks 46 to 51 (P < 0.05). No significant differences were found between the control and FG group. There was no obvious difference on plasma levels of total protein, blood urea nitrogen, uric acid, and activities of albumen, alanine aminotransferase, aspartate aminotransferase and the histopathology of liver, kidney, and oviduct in all groups. In conclusion, decreasing feed intake, egg weight, and egg quality were observed in the 189 g/kg LCSM group. However, the adverse effect was not relevant to the presence of FG in LCSM. Dietary LCSM supplementation of 98.3 g/kg was recommended with optimum FCR and without adverse effect on egg production, quality, and health of layers.

Bioefficacy comparison of organic manganese with inorganic manganese for eggshell quality in Hy-Line Brown laying hens.

This study was aimed at investigating the bioefficacy of organic compared with inorganic manganese (Mn) for eggshell quality. An amino acid-Mn complex or Mn sulfate monohydrate was used as the organic or inorganic Mn source. A total of six hundred forty-eight 50-wk-old layers (Hy-Line Brown) were divided into 9 groups; each group consisted of 6 replicates with 12 layers each. The feeding trial lasted 12 wk. During the first 4 wk of the feeding trial, the groups were fed a basal diet, which met the nutrient requirements of the layers, except for Mn. During the following 8 wk, 9 levels of Mn (inorganic Mn: 0, 25, 50, 100, and 200 mg/kg; organic Mn: 25, 50, 100, and 200 mg/kg) were used to supplement, respectively, in the basal diet on an equimolar basis. An exponential regression model was applied to calculate the bioefficacy of organic Mn compared with the inorganic Mn. Dietary supplementation with either organic or inorganic Mn did not influence egg production and feed efficiency of (P > 0.05), and eggshell quality did not exhibit a significant response to dietary supplementation with Mn sources at 56 and 58 wk (P > 0.05). Dietary supplementation with either organic Mn or inorganic Mn significantly enhanced the thickness, breaking strength, and elastic modulus of the eggshells compared with the control group at the end of 62 wk (P < 0.05). At the end of 62 wk, the bioefficacy of organic Mn was 357% (shell thickness), 406% (breaking strength), 458% (elastic modulus), and 470% (eggshell Mn), as efficacious as inorganic Mn at equimolar levels. This study suggests that organic Mn enhances eggshell quality in aged laying hens compared with inorganic Mn.

Effects of dietary pyrroloquinoline quinone disodium on growth, carcass characteristics, redox status, and mitochondria metabolism in broilers.

The potential benefits of supplementing pyrroloquinoline quinone disodium (PQQ·Na2) in the diet of broiler chicks were explored. We first examined the effect of different levels of dietary PQQ·Na2 on growth performance, carcass characteristics, and plasma biochemical parameters (trial 1). A total of 490 1-day-old male Arbor Acres (AA) broiler chicks were randomly divided into 5 dietary groups supplemented with 0, 0.05, 0.1, 0.2, or 0.4 mg PQQ·Na2/kg feed. As the 0.2 mg/kg PQQ·Na2 supplement gave the best performance, we then investigated whether this level of PQQ·Na2 influenced the redox status of plasma samples and mitochondrial-related metabolism (trial 2). A total of 120 1-day-old male AA chicks were randomly divided into 2 groups supplemented with 0 or 0.2 mg PQQ·Na2/kg diet. In trial 1, birds fed a diet containing 0.2 mg PQQ·Na2/kg showed lower feed conversion ratio compared with those fed the control diet in the overall study (d 1 to 42, P=0.039). Breast muscle yield (d 42) increased quadratically in response to dietary PQQ·Na2 supplementation (P=0.021). Analysis of plasma biochemical parameters revealed that feeding broiler chicks with ≤0.4 mg/kg PQQ·Na2 did not cause adverse health effects. In trial 2, birds fed 0.2 mg/kg PQQ·Na2 again showed improved feed efficiency than the control birds in the grower and overall phases (P=0.038 and 0.016, respectively). In addition, dietary PQQ·Na2 supplementation resulted in a higher anti-oxidative capacity (P=0.001), lower redox potential (P=0.008), and higher hepatic citrate synthase activity (P=0.002). In contrast, no difference in hepatic mitochondrial DNA copy number was observed between the 2 experimental groups (P>0.1). These results indicate that PQQ·Na2 is a potentially effective feed additive for improving feed efficiency, stimulating breast muscle development, and maintaining redox status in broiler chicks. Enhancement of mitochondria efficiency, rather than modulating mitochondria numbers, may underlie the growth-promoting effect of PQQ·Na2.

Effects of dietary pyrroloquinoline quinone disodium on growth performance, carcass yield and antioxidant status of broiler chicks.

Pyrroloquinoline quinone (PQQ), a putative essential nutrient and redox modulator in microorganisms, cell and animal models, has been recognized as a growth promoter in rodents. Growth performance, carcass yield and antioxidant status were evaluated on broiler chickens fed different levels of PQQ disodium (PQQ.Na2). A total of 784 day-old male Arbor Acres (AA) broilers were randomly allotted into seven dietary groups: negative control group (NC) fed a basal diet without virginiamycin (VIR) or PQQ.Na2; a positive control group (PC) fed a diet with 15 mg of VIR/kg diet; and PQQ.Na2 groups fed with 0.05, 0.10, 0.20, 0.40 or 0.80 mg PQQ.Na2/kg diet. Each treatment contained eight replicates with 14 birds each. The feeding trial lasted for 6 weeks. The results showed that chicks fed 0.2 mg PQQ.Na2/kg diet significantly improved growth performance comparable to those in PC group, and the feed efficiency enhancement effects of dietary PQQ.Na2 was more apparent in grower phase. Dietary addition of PQQ.Na2 had the potential to stimulate immune organs development, and low level dietary addition (<0.1 mg/kg) increased plasma lysozyme level. Broilers fed 0.2 mg PQQ.Na2/kg diet gained more carcasses at day 42, and had lower lipid peroxide malondialdehyde content and higher total antioxidant power in plasma. The results indicated that dietary PQQ.Na2 (0.2 mg/kg diet) had the potential to act as a growth promoter comparable to antibiotic in broiler chicks.

Manganese supplementation enhances the synthesis of glycosaminoglycan in eggshell membrane: a strategy to improve eggshell quality in laying hens.

This study investigated the effect of dietary Mn supplementation on eggshell quality, ultrastructure, glycosaminoglycan (GAG), and uronic acid content, and mRNA and protein expression of Galß1,3-glucuronosyltransferase (GlcAT-I). A total of 216 layers (Hy-Line Grey) at age of 50 wk were divided into 3 groups. In the first 8 wk of the 12-wk feeding trial, all groups were fed a basal diet that met all layer nutrient requirements except for Mn. In the last 4 wk, each group was fed 1 of 3 diets supplemented with Mn levels at 0, 25, or 100 mg Mn/kg. Dietary Mn deficiency did not affect the egg performance of layers. Dietary Mn supplementation significantly improved the breaking strength, thickness, and fracture toughness of eggshells (P < 0.05). In photographs of eggshell ultrastructure, the size of mammillary cones and cracks in the outer surface were decreased by dietary Mn supplementation. The contents of GAG and uronic acids in eggshell membrane were significantly increased by dietary Mn addition (P < 0.05). This result was further confirmed by increased mRNA expression and protein expression of GlcAT-I when Mn was added to the diet. This study suggests that dietary Mn supplementation can improve eggshell quality by enhancing the GAG and uronic acid synthesis in the eggshell glands, which can affect the ultrastructure of eggshells.

Stimulation with monochromatic green light during incubation alters satellite cell mitotic activity and gene expression in relation to embryonic and posthatch muscle growth of broiler chickens.

Previous studies showed that monochromatic green light stimuli during embryogenesis accelerated posthatch body weight (BW) and pectoral muscle growth of broilers. In this experiment, we further investigated the morphological and molecular basis of this phenomenon. Fertile broiler eggs (Arbor Acres, n=880) were pre-weighed and randomly assigned to 1 of the 2 incubation treatment groups: (1) dark condition (control group), and (2) monochromatic green light group (560 nm). The monochromatic lighting systems sourced from light-emitting diode lamps and were equalized at the intensity of 15 lx at eggshell level. The dark condition was set as a commercial control from day 1 until hatching. After hatch, 120 male 1-day-old chicks from each group were housed under incandescent white light with an intensity of 30 lx at bird-head level. No effects of light stimuli during embryogenesis on hatching time, hatchability, hatching weight and bird mortality during the feeding trial period were observed in the present study. Compared with the dark condition, the BW, pectoral muscle weight and myofiber cross-sectional areas were significantly greater on 7-day-old chicks incubated under green light. Green light also increased the satellite cell mitotic activity of pectoral muscle on 1- and 3-day-old birds. In addition, green light upregulated MyoD, myogenin and myostatin mRNA expression in late embryos and/ or newly hatched chicks. These data suggest that stimulation with monochromatic green light during incubation promote muscle growth by enhancing proliferation and differentiation of satellite cells in late embryonic and newly hatched stages. Higher expression of myostatin may ultimately help prevent excessive proliferation and differentiation of satellite cells in birds incubated under green light.

Solexa sequencing of microRNAs on chromium metabolism in broiler chicks.

AIM: The aim of this study was to determine the effect of chromium picolinate (CrPic) on the differential expression of the known microRNAs (miRNAs) in broiler skeletal muscle. METHODS AND RESULTS: A total of 288 1-day-old male Arbor Acres broilers were randomly assigned to one of four dietary treatments supplemented with 0, 0.4, 2.0, or 10.0 mg·kg(-1) CrPic, respectively. Dietary CrPic supplementation at 10.0 mg·kg(-1) increased the average daily feed intake in broilers (p < 0.05). On day 42, the serum total protein level was highest in animals treated with 2.0 mg·kg(-1) (p < 0.05) and 10.0 mg·kg(-1) CrPic (p < 0.05). Dietary supplementation with 10.0 mg·kg(-1) CrPic decreased the levels of serum glucose (p < 0.05) on day 42 and of serum triglyceride (p < 0.05) on days 21 and 42. To further identify miRNAs from broiler skeletal muscles, we sequenced two small RNA libraries using the Solexa sequencing approach, and 57 miRNAs were found to be significantly differentially expressed (p < 0.05). Among them, 6 upregulated and 2 downregulated miRNAs were validated by real-time qPCR (p < 0.05). CONCLUSIONS: The results of the present study provide a valuable clue regarding the role of miRNA target genes in the mechanism of the dietary CrPic effect on protein synthesis in skeletal muscles of broilers.

Trimethylamine deposition in the egg yolk from laying hens with different FMO3 genotypes.

The A/T polymorphism at position nt 1,034 of the chicken FMO3 cDNA sequence is associated with elevated levels of trimethylamine (TMA) in the egg yolk, which is responsible for the fishy egg flavor. This study was aimed to investigate yolk TMA deposition in eggs from different FMO3 genotype (AA, AT, TT) laying hens fed a high level of choline, and the relationship between egg yolk TMA contents and yolk acceptability. A total of 132 genotyped laying hens, 38 wk of age, were assigned to 1 of the 4 dietary treatments: 1) a control group of AA hens fed 370 mg of choline/kg of diet (practical choline level, provided by 500 mg of choline chloride/kg of diet); 2) AA hens were fed 2,960 mg of choline/kg of diet (higher dietary choline level, provided by 4,000 mg of choline chloride/kg of diet); 3) AT hens were fed 2,960 mg of choline/kg of diet; and 4) TT hens were fed 2,960 mg of choline/kg of diet. All layers were fed a 370 mg of choline/kg added diet for a period of 1-wk adaptation followed by a 6-wk trial period. A remarkable increase in yolk TMA concentration of TT hens (P < 0.001) caused by dietary choline addition at 2,960 mg/kg was observed. The relationship between TMA concentration in the egg yolk (µg/g, y) and deposition time (1~42 d, x) for TT hens is y = 0.0005x(3) - 0.0419x(2) + 1.0924x + 0.4323 (R(2) = 0.9259). The fish-flavor scores of egg yolks rose steadily (R(2) = 0.9324) as the TMA concentration increased, and there was also a corresponding decrease in the acceptance score (R(2) = 0.8276). The eggs were acceptable when the yolk TMA concentrations were less than 4.516 µg/g of yolk.

Effect of ß-hydroxy-ß-methylbutyrate calcium on growth, blood parameters, and carcass qualities of broiler chickens.

Beta-hydroxy-ß-methylbutyrate (HMB), the metabolite of leucine, plays an important role in muscle protein metabolism. To investigate the effect of dietary HMB calcium (HMB-Ca) on growth performance, breast muscle development, and serum parameters in broiler chickens, a total of two hundred seventy 1-d-old Arbor Acres male broiler chicks were randomly allotted into 3 dietary treatments supplemented with 0, 0.05%, or 0.1% HMB-Ca during the starter (1 to 21 d) and grower (22 to 42 d) period. The results showed that broilers fed 0.1% HMB-Ca diet had higher ADG during the starter or the whole period, and gain 148 g more BW than the chicks fed the control diet at 42 d of age (P < 0.05). At 21 d of age, birds receiving 0.1% HMB-Ca had more breast muscle yield, less abdominal fat than the control, and more dressing percentage than birds fed the control or 0.05% HMB-Ca diet (P < 0.05). At 42 d of age, 0.1% HMB-Ca increased breast muscle yield than the control and decreased abdominal fat compared with the control or 0.05% HMB-Ca group (P < 0.05). In comparison with the control, feeding 0.1% HMB-Ca increased the triiodothyronine, thyroxine, triiodothyronine/thyroxine ratio and decreased the serum uric acid level at d 21 (P < 0.05). At 42 d of age, serum thyroxine level was elevated in the 0.05% HMB-Ca treatment, and the uric acid concentration was significantly decreased by the 0.1% HMB-Ca-supplemented diet (P < 0.05). Dietary HMB-Ca did not affect the growth hormone or insulin content. This study suggested that dietary supplementation of HMB-Ca improved growth performance, stimulated the breast muscle development, and decreased the abdominal fat deposition in broiler chickens, and the favorable effects were more pronounced in the starter phase. The growth promotion effect of HMB-Ca may be partly related to the increased serum thyroid hormones in broiler chickens.

Effect of dietary choline supplementation under different flavin-containing monooxygenase 3 genotypes on trimethylamine metabolism in laying hens.

To evaluate the effect of flavin-containing monooxygenase 3 (FMO3) genotype and dietary choline supplementation on trimethylamine (TMA) metabolism in HyLine Brown laying hens, a 3 × 2 two-factorial arrangement was employed with FMO3 genotypes (AA, AT, and TT) and dietary choline supplemental levels (370 and 2,960 mg/kg of diet) as main effects. At 46 wk of age, 108 hens of AT genotype and 108 hens of TT genotype were randomly allotted to one of the 2 dietary treatments, and each dietary treatment consisted of 6 replicates with 9 birds each. A total of 24 hens with AA genotype was allotted to one of the 2 dietary treatments that consisted of 6 replicates with 2 hens. Hens were fed the diet with 370 mg/kg of choline supplementation for 1 wk of adaptation followed by a 6-wk trial period. Yolk TMA concentration was increased by dietary supplemental choline at 2,960 mg/kg (P < 0.05), and TT hens showed a higher TMA content in egg yolks than that in AA and AT hens (P < 0.05). Dietary supplementation of choline at 2,960 mg/kg increased the TMA concentration of cecal chyme (P < 0.05) and serum (P < 0.05). Hepatic FMO3 mRNA levels in hens were reduced by higher choline added to the diet (P < 0.05). The TMA and methimazole oxidation rate in AA hens was higher than those in AT and TT hens (P < 0.05). A higher choline diet decreased hepatic FMO3 activity by 33.99% (P < 0.05) and 61.39% (P < 0.05) toward TMA and methimazole, respectively. These results suggest that lower hepatic FMO3 activity caused by the mutation may be responsible for the genotype difference in the TMA metabolism. Exposure to a high dosage of dietary choline increased TMA synthesis in the cecum, suppressed activity of FMO3 in liver, and consequently aggravated the burden of TMA metabolism, especially in TT hens.

Effect of monochromatic light stimuli during embryogenesis on muscular growth, chemical composition, and meat quality of breast muscle in male broilers.

This study was conducted to evaluate the effect of monochromatic light stimuli during embryogenesis on breast muscle growth, chemical composition, and meat quality of male broilers. Fertile broiler eggs (Arbor Acres; n = 1,320) were preweighed and randomly assigned to 1 of 3 treatment groups in 3 modified incubators: 1) control group (in dark condition), 2) monochromatic green light group (560 nm), and 3) monochromatic blue light group (480 nm). The monochromatic lighting systems sourced from light-emitting diode lamps and were equalized at the intensity of 15 lx at eggshell level. After hatch, 120 male chicks from each group were placed in 6 replicates with 20 birds each. All of the birds were housed under white light (30 lx at bird-head level) with a light schedule of 23L:1D. At 21, 35, and 42 d of age, BW and breast muscle weight in the green light group were significantly increased compared with birds in the blue or dark groups (P < 0.05). The breast muscle weight and breast muscle percentages in birds incubated under green light were significantly elevated by 50.39 g (0.76%) and 54.07 g (1.20%) than those in the dark condition or blue group at 42 d of market age (P < 0.05), respectively. In the green light group, feed intake during 0~42 d was higher than that in the other 2 treatment groups (P < 0.05); feed conversion ratio during 0~35 and 0~42 d were lower than that in the dark condition (P < 0.05). No significant differences in the contents of breast moisture, CP, crude fat, and crude ash among all groups were observed (P > 0.05). Green light stimuli tended to increase cooking loss (P = 0.08) and L* value of 24-h meat color (P = 0.09). These results suggest that green light stimuli during embryogenesis enhanced the posthatch BW of male broilers, increased breast muscle growth, and improved the feed conversion ratio, but it did not cause any noticeable changes in breast chemical composition or overall meat quality characteristics.

Comparison of lipid oxidation, messenger ribonucleic acid levels of avian uncoupling protein, avian adenine nucleotide translocator, and avian peroxisome proliferator-activated receptor-γ coactivator-1α in skeletal muscles from electrical- and gas-stunned broilers.

The aim of this study was to compare the effects of stunning methods [electrical stunning (ES) vs. gas stunning (GS)] on lipid oxidation in broiler meat and to investigate possible mechanisms of lipid oxidation by measuring plasma variables, muscle reactive oxygen species (ROS), and TBA reactive substance (TBARS) concentrations, muscle fiber ratios, and mRNA levels of avian uncoupling protein (avUCP), avian adenine nucleotide translocator, and avian peroxisome proliferator-activated receptor-γ coactivator-1α (avPGC-1α). Arbor Acres broilers (n = 36) were not stunned (control) or were exposed to the following stunning treatments: 40% CO(2) + 21% O(2) + N(2); 60% CO(2) + 21% O(2) + N(2); 35 V, 47 mA, 400 Hz; 50 V, 67 mA, 160 Hz; and 65 V, 86 mA, 1,000 Hz. The ROS level in tibialis anterior (TA; P < 0.05) and the TBARS concentration in pectoralis major (PM; P < 0.01) were decreased in the GS groups compared with the ES groups at 45 min postmortem. However, the TBARS concentrations at 24 h postmortem in the PM and TA groups were not affected by stunning method (ES or GS). Compared with ES, GS caused greater expression of avUCP mRNA (1.47-fold in PM, and 2.41-fold in TA) and avPGC-1α mRNA (1.42-fold in PM, and 2.08-fold in TA). In conclusion, the upregulation of avUCP and avPGC-1α reduced ROS accumulation and lipid oxidation at 45 min postmortem in the skeletal muscles of broilers stunned with hypercapnic moderate oxygenation GS. However, these changes were not sufficient to cause a difference in meat lipid oxidation at 24 h postmortem between broilers stunned with hypercapnic moderate oxygenation GS and those stunned with low-current, high-frequency ES.