Cambridge University Press
Supplemental Table S1. Composition of riboflavin-deficient basal diet (as-fed basis). ItemBasal diet (from hatch to day 21)Ingredient, g/kgCorn622.4Soybean120.0Corn gluten meal210.0Limestone10.0Dicalcium phosphate16.0Vitamin and trace mineral premix*10.0Sodium chloride3.0DL-Methionine 0.7L-Lysine·HCl5.3L-Threonine0.6L-Tryptophan1.0L-Arginine1.0Calculated compositionMetabolizable energy?, MJ/kg 12.64Crude protein, g/kg200.0Calcium, g/kg9.9Nonphytate phosphorus, g/kg4.2Lysine, g/kg 11.0Methionine, g/kg4.7Methionine + cysteine, g/kg 8.2Threonine, g/kg 7.6Tryptophan, g/kg 2.2Arginine, g/kg 10.0Riboflavin?, mg/kg 1.38* Supplied per kilogram of total diet: Cu (CuSO4?5H2O), 10 mg; Fe (FeSO4?7H2O), 60 mg; Zn (ZnO), 60 mg; Mn (MnSO4?H2O), 80 mg; Se (NaSeO3), 0.3 mg; I (KI), 0.2 mg; choline chloride, 1000 mg; vitamin A (retinyl acetate), 10000 IU; vitamin D3 (Cholcalciferol), 3000 IU; vitamin E (DL-α-tocopheryl acetate), 20 IU; vitamin K3 (menadione sodium bisulfate), 2 mg; thiamin (thiamin mononitrate), 2 mg; pyridoxine hydrochloride, 4 mg; cobalamin, 0.02 mg; calcium-D-pantothenate, 20 mg; nicotinic acid, 50 mg; folic acid, 1 mg; biotin, 0.2 mg.? The values are calculated according to the AME of chickens (Ministry of Agriculture of China, 2004).? The amount was determined using high performance liquid chromatography.Supplemental Table S2. Growth performance on day 21 of ducks in the riboflavin-deficient (RD), pair-fed (CPF), and ad. libitum control (CAL) groups from hatch to 21 days of ageVariable RDCPFCALSEMP-valueMortality (%)9.38a0.00b 1.04b-0.002ADG (g/d)15.1c17.2b60.7a3.57<0.001ADFI (g/d)29.4b29.1b100.3a5.71<0.001G:F / (g:g)0.51b0.59a0.61a0.008<0.001Relative liver weight *4.47a3.51b3.36b0.15<0.001ADG, average daily weight gain; ADFI, average daily feed intake; G:F, gain to feed ratio; and SEM, standard error of the mean. a, b, c Mean values with unlike superscript letters were significantly different (P < 0.05). Data were analyzed by one-way ANOVA followed by Tukey’s multiple comparison. Data was expressed as mean and SEM (n=12). * Relative liver weight were calculated as liver weight:body weight (g/100 g).Supplemental Table S3. Liver fatty acid composition (% of the total fatty acid) on day 21 of ducks in the riboflavin-deficient (RD), pair-fed (CPF), and ad. libitum control (CAL) groups% of the total fatty acidRD CPFCALSEMP-valueC6:00.133a0.090b0.048c0.0120.001C8:00.013a0.013a0.010b0.0010.026C10:00.0200.0210.0190.0010.755C12:00.041a0.032b0.029b0.0020.016C14:00.372b0.472a0.403b0.0170.025C14:10.044b0.069a0.049b0.0040.001C15:00.0410.0380.0220.0040.052C16:025.2a23.9b21.5c0.472<0.001C16:11.14b1.07b1.40a0.0590.037C17:00.116a0.125a0.094b0.0050.011C18:024.7a21.2b17.3c1.010.001C18:1n9c22.3b19.2b29.7a1.640.008C18:2n6c6.90b9.98a7.31b0.431<0.001C18:3n30.1390.1360.0990.0210.732C20:00.222a0.227a0.175b0.0080.003C20:10.384b0.310c0.453a0.0210.002C21:01.08b1.33b2.19a0.151<0.001C20:20.031a0.030a0.021b0.0020.029C20:3n61.00b1.51a1.40a0.0830.012C20:4n613.717.315.20.6790.082C20:3n30.0520.0390.0470.0050.653C20:5n30.076b0.121a0.135a0.0090.002C22:00.376b0.514a0.345b0.0240.0003C22:1n90.0400.0440.0510.0030.2492C23:00.243b0.392a0.263b0.0230.001C24:00.490b0.765a0.512b0.041<0.001C22:6n30.7420.6590.6640.0400.679C24:10.4840.4840.4730.0180.964Total SFA53.0a49.1b43.0c1.35<0.001Total MUFA24.4b21.2b32.1a1.690.007Total PUFA22.7b29.8a24.9b1.140.013Total n-31.010.950.950.050.891Total n-621.6b28.8a23.9b1.120.008n-6:n-322.3b30.5a25.5ab1.300.016SFA, saturated fatty acid; MUFA, mono-unsaturated fatty acid; PUFA, poly-unsaturated fatty acid.a, b, c Mean values with unlike superscript letters were significantly different (P < 0.05). Data were analyzed by one-way ANOVA followed by Tukey’s multiple comparison. Data was expressed as mean and SEM (n=12).Supplemental Table S4. Differentially expressed proteins in liver on day 21 of ducks caused by riboflavin deficiency (RD)Accession no.Protein nameGene nameFold change*RD vs. CALRD vs. CPFCPF vs. CALtr|R7VUZ3|R7VUZ3_COLLIFrataxinFXN-90.9-90.9-1.05tr|Q9DG15|Q9DG15_CHICKP95-APP1none-90.9-90.9-1.10tr|R7VXW2|R7VXW2_COLLIInositol polyphosphate 1-phosphataseINPP1-83.3-90.92.33tr|Q38HW8|Q38HW8_ANAPLThyroid hormone responsive Spot 14 alphaTHRSP-62.5-71.41.24tr|U3JV22|U3JV22_FICALFlotillin-2FLOT2-62.5-71.41.27tr|F1NAM7|F1NAM7_CHICK3-hydroxybutyrate dehydrogenase type 2BDH2-23.8-23.31.43tr|U3I9X5|U3I9X5_ANAPLAnnexinANXA7-16.4-76.94.66tr|U3J4D8|U3J4D8_ANAPLPrefoldin subunit 3VBP1-14.1-14.7-1.41tr|U3I0A3|U3I0A3_ANAPLKeratin, type II cytoskeletal 7KRT7-11.6-8.85-1.47tr|R0K082|R0K082_ANAPLNADH-ubiquinone oxidoreductase 75 kDa subunitNDUFS1-11.4-11.11.15RRRRRtr|F1NSA7|F1NSA7_CHICKNone LOC100857563-9.62-9.62-1.16tr|Q8AYP9|Q8AYP9_CHICK6-phosphofructokinasePFK-8.00-9.261.13tr|U3I7F8|U3I7F8_ANAPL39S ribosomal protein L21MRPL21-7.58-6.371.21tr|U3J1J0|U3J1J0_ANAPLAcyl-CoA dehydrogenase family member 9ACAD9-6.41-8.401.41tr|U3IL38|U3IL38_ANAPLMalic enzymeME1-6.02-8.701.36tr|U3IR48|U3IR48_ANAPLDihydrolipoyl dehydrogenaseDLD-5.71-8.261.47tr|U3J8W0|U3J8W0_ANAPLShort-chain specific acyl-CoA dehydrogenaseACADS-4.74-6.801.17tr|Q5ZJZ9|Q5ZJZ9_CHICKProtein arginine N-methyltransferase 5PRMT5-4.69-21.34.83tr|U3IHZ7|U3IHZ7_ANAPLDelta-aminolevulinic acid dehydrataseALAD-3.83-4.171.09tr|U3ITA9|U3ITA9_ANAPLMedium-chain specific acyl-CoA dehydrogenaseACADM-2.99-3.251.08tr|U3J3V1|U3J3V1_ANAPLElectron transfer flavoprotein-ubiquinone oxidoreductaseETFDH-2.88-3.191.06tr|U3JW36|U3JW36_FICALCoactosin-like F-actin binding protein 1COTL1-2.83-7.813.08tr|U3J680|U3J680_ANAPLCaldesmonCALD1-2.75-4.371.33tr|G1N071|G1N071_MELGAMalate dehydrogenaseMDH1-1.98-2.27-1.01tr|E1C958|E1C958_CHICKLegumainLGMN-1.98-1.75-1.13tr|U3IQB4|U3IQB4_ANAPLHydroxyacid oxidase 2HAO2-1.92-1.60-1.21tr|U3J788|U3J788_ANAPLAlveolar soft part sarcoma chromosome region, candidate 1ASPSCR1-1.68-2.031.20sp|Q5ZML3|SRSF1_CHICKSerine/arginine-rich splicing factor 1SRSF1-1.68-1.821.08tr|U3ID88|U3ID88_ANAPLCollagen alpha-3(VI) chainCOL6A3-1.66-2.131.25tr|U3J5K2|U3J5K2_ANAPLNADH dehydrogenase [ubiquinone] 1 alpha subcomplex subunit 8NDUFA8-1.64-3.402.03tr|U3IAF7|U3IAF7_ANAPLCopine-1CPNE1-1.63-1.711.12tr|U3INW1|U3INW1_ANAPLSideroflexin-1SFXN11.511.94-1.32tr|U3IJZ6|U3IJZ6_ANAPLFatty acid amide hydrolaseFAAH1.592.17-1.36tr|U3IL01|U3IL01_ANAPLHeme oxygenase 1HMOX11.681.94-1.22tr|U3J6P0|U3J6P0_ANAPLComplement component 3C31.796.86-3.83tr|U3IXZ9|U3IXZ9_ANAPL4-aminobutyrate aminotransferaseABAT1.792.54-1.46tr|U3J8L3|U3J8L3_ANAPLSolute carrier family 25 (aspartate/glutamate carrier), member 13SLC25A131.844.88-2.81tr|U3IKU2|U3IKU2_ANAPLNon-specific lipid-transfer proteinSCP21.922.23-1.13tr|U3IEH8|U3IEH8_ANAPLAcetyl-CoA acetyltransferaseACAT22.001.601.26tr|U3IWA4|U3IWA4_ANAPLApolipoprotein B-100APOB2.012.97-1.45tr|U3IZY1|U3IZY1_ANAPLAldehyde dehydrogenase 7 family, member A1ALDH7A12.133.95-1.94tr|U3I3S5|U3I3S5_ANAPLNicotinamide nucleotide transhydrogenaseNNT2.214.41-2.17tr|U3IU70|U3IU70_ANAPLXanthine dehydrogenase/oxidaseXDH2.272.071.10tr|R0K9V0|R0K9V0_ANAPLAcetyl-coenzyme A synthetaseACS2.314.61-2.13tr|R0JGG2|R0JGG2_ANAPLHydroxymethylglutaryl-CoA synthase, cytoplasmicHMGCS12.424.21-1.82tr|R0JWK3|R0JWK3_ANAPLLipoamide acyltransferase component of branched-chain alpha-keto acid dehydrogenase complexDBT2.421.601.41tr|F1NFC6|F1NFC6_CHICK40S ribosomal protein S4, X isoformRPS4X2.473.73-1.54tr|U3III1|U3III1_ANAPLAcyl-coenzyme A thioesterase 1ACOT12.614.17-1.66tr|U3I2B5|U3I2B5_ANAPLSolute carrier family 27 (fatty acid transporter), member 5SLC27A52.755.45-2.29tr|U3IE74|U3IE74_ANAPLL-lactate dehydrogenaseLDHA2.8111.7-5.85tr|U3J3G1|U3J3G1_ANAPLEnoyl-CoA hydrataseECHS12.832.781.05tr|R0JLG5|R0JLG5_ANAPLCalnexinCANX3.136.61-2.15tr|U3I559|U3I559_ANAPLMicrosomal triglyceride transfer proteinMTTP3.229.20-3.40tr|U3IU30|U3IU30_ANAPLLong-chain-fatty-acid--CoA ligase 1ACSL13.374.17-1.20tr|U3J4Z9|U3J4Z9_ANAPLLong-chain-fatty-acid--CoA ligase 5ACSL53.579.73-3.25tr|U3J7S6|U3J7S6_ANAPLNone LOC1017965964.173.601.09tr|U3J4X1|U3J4X1_ANAPLHeat shock protein 90kDa beta (Grp94), member 1HSP90B14.663.631.21tr|R0LYJ5|R0LYJ5_ANAPLTrifunctional purine biosynthetic protein adenosine-3GART5.064.371.13tr|R0KBX6|R0KBX6_ANAPLProtein disulfide-isomerasePDIA48.2410.1-1.02tr|H0ZD22|H0ZD22_TAEGU39S ribosomal protein L38MRPL3871.871.8-1.01RRRRRtr|R4GKG1|R4GKG1_CHICKTANK-binding kinase 1-binding protein 1TBKBP180.980.9-1.01tr|H0ZF88|H0ZF88_TAEGUAcyl-CoA synthetase family member 2ACSF286.386.3-1.01RRRRRtr|U3IQK5|U3IQK5_ANAPLSki oncogeneSKI87.987.9-1.01CPF: pair-fed; CAL: ad. libitum control* Fold change are expressed as the ratio between two of three groups. For diminished proteins, the fold change was transformed to the corresponding negative value.Supplemental Table S5. Enriched GO (biological processes)-based sets of differentially expressed proteins in the liver of starter Pekin ducks in response to riboflavin deficiency GO termGO accessionP valueCountLipid metabolism fatty acid metabolic processGO:000663117.0E-1517fatty acid oxidationGO:00193952.7E-98acylglycerol metabolic processGO:00066398.3E-99cellular lipid catabolic processGO:004424228.0E-99lipid modificationGO:003025849.0E-99acylglycerol biosynthetic processGO:0046463750.0E-96long-chain fatty acid metabolic processGO:00016761.8E-66fatty-acyl-CoA metabolic processGO:003533731.0E-64Cellular respirationcellular respirationGO:004533362.0E-99respiratory electron transport chainGO:00229049.8E-66aerobic respirationGO:0009060110.0E-64oxidative phosphorylationGO:0006119360.0E-64mitochondrial electron transport, NADH to ubiquinoneGO:0006120610.0E-63mitochondrial respiratory chain complex I assemblyGO:00329811.0E-33Other processescarboxylic acid catabolic processGO:004639569.0E-1212cofactor biosynthetic processGO:0051188360.0E-1210coenzyme biosynthetic processGO:0009108630.0E-97heme metabolic processGO:00421684.7E-64Supplemental Fig. S1. Plasma riboflavin concentrations (A) and liver riboflavin, flavin mononucleotide (FMN), and flavin adenine dinucleotide (FAD) concentrations (B) in 21-day-old ducks from the riboflavin-deficient (RD), pair-fed (CPF), and ad. libitum control (CAL) groups, respectively. Data are means ± SEM (n=12). Mean values with unlike superscript letters were significantly different (P < 0.05). Data were analyzed by one-way ANOVA followed by Tukey’s multiple comparison. ................
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