<p>Suppl. Table 1 Prepartum diet for the dry off cows Close-up Item diet (CUD) Ingredient % of DM Alfalfa hay 10.0 Barley silage 60.0 CUD grain 30.0 Nutrient composition of CUD grain % in 100 kg of mix Ruminant TM Pak1 0.2775 Selenium 1000 mg/kg (UNscr 0.2 FineCr) Custom TM Complex Premix2 0.33 3 Vitamin A/D3-1000-200 0.006 Barley grain, rolled 39.5815 Flo-bond mycotoxin binder 0.5 Limestone 3.7 Magnesium chloride 1.645 Mag Ox-56%4 0.54 Scale Molasses (60:40) 2.5 Canola meal 17.0 Vitamin E 50% Ads5 0.18 Soybean hulls, ground 33.0 Salt 0.54 1Ruminant TM Pak: a premix containing cobalt, copper, iodine, manganese, and zinc. 2Custom TM complex premix: a custom product supplying organic sources of cobalt, copper, manganese, and zinc. 3 Vitamin A/D3-1000-2003: Vitamin A acetate (retinyl acetate) and Vitamin D3 (cholecalciferol). 4Mag Ox 56%: 56% magnesium guarantee. 5Vitamin E 50% Ads contains 226,800 IU of Vitamin E per pound. Suppl. Table 2 Ingredients of TMR fed to cows during early lactation Early Item lactation diet Ingredient % of DM %, DM Alfalfa Hay 9.59 Barley Silage 30.24 Alfalfa Silage 9.64 High 16% dairy ration 50.53 Nutrient composition of dairy ration % amount per kg ADE Vit Pak-30 Natural E1 0.05 Ruminant TM Pak2 0.11 Selenium, 1,000 mg/kg (UNscr 0.07 FineCr) Custom TM Complex premix3 0.07 AminoShure - L4 0.33 Blood meal 3.50 Barley grain, rolled 39.90 Barley grain, ground 27.50 Di-calcium phosphate 21% 1.00 Vit D-10,000 KIU/kg 0.02 Diamond V XPC5 0.13 Dairy Xtract 0.02 Energizer RP10 2.75 Limestone 1.70 Mag Ox-56%6 0.43 Scale Molasses (60:40) 1.25 Nutri A-Z C Dry 0.10 Amino Plus (High bypass soy)7 8.00 Vitamin E 50% Ads8 0.01 Soy bean meal-47.5% 1.25 Sodium bicarbonate 0.80 Salt 0.50 Poultry-Tallow 0.50 Biotin 2%-Rovimix H-29 0.01 Wheat distillers grain (50:50) 10.00 1ADE Vit Pak-30 Natural E: a premix containing vitamins A, D3, and E. 2Ruminant TM Pak: a premix containing cobalt, copper, iodine, manganese, and zinc. 3Custom TM complex premix: a custom product supplying organic sources of cobalt, copper, manganese, and zinc. 4AminoShure - L: hydrogenated vegetable oil, and L-lysine monohydrochloride (Halchemix, Port Perry, ON, Canada). 5Diamond V XPC: concentrated yeast (Diamond V Mills, Cedar Rapids, IA). 6Mag Ox 56%: 56% magnesium guarantee. 7Amino Plus: a high by-pass soy meal. 8Vitamin E 50% Ads contains 226,800 IU of Vitamin E per pound. 9DSM Nutritional Products (Parsippany, NJ).</p><p>Suppl. Table 3 Concentrations of serum metabolites (mean (SEM)) in healthy control (CON) and ketosis cows at 3 time points (-8 wk, -4 wk, and the wk of diagnosis of disease) as determined by DI/LC-MS/MS</p><p>4 week before parturition Ketosis diagnosis week1 C Fold Ketosis / O Ketosis / Ketosis / Metabolite, μM2 CON P-value change CON Ketosis N P-value Fold change CON Ketosis CON P-value Fold change CON </p><p>Number of cases 20 - - - 6 20 - - - 6 20 - - - 0. 03 (0. 0.02 00 C16 (0.003) 1 1 Down 0.01 (0.003) 3) 0.447 -3 Down 0.05 (0.01) 0.02 (0.00) 0.003* 2.5 Up 0. 04 (0. 0.03 01 C18 (0.003) 0.519 -1.5 Down 0.02 (0.004) ) 0.192 -2 Down 0.09 (0.02) 0.04 (0.01) 0.014* 2.25 Up 1. 43 (0. 1.30 22 C2 (0.10) 0.869 1.14 Up 1.17 (0.08) ) 0.717 -1.22 Down 2.59 (0.47) 1.22 (0.11) 0.004* 2.12 Up 0. 14 (0. 0.14 02 C3 (0.01) 0.488 1 Up 0.14 (0.02) ) 0.621 1 Up 0.13 (0.02) 0.15 (0.05) 0.219 -1.15 Down 0. 07 (0. 0.08 01 C4 (0.01) 0.717 1.13 Up 0.08 (0.01) ) 0.476 1.14 Up 0.08 (0.01) 0.07 (0.00) 0.242 1.14 Up 0. 06 (0. 0.08 01 C5 (0.01) 0.519 -1.14 Down 0.06 (0.004) ) 0.974 1 Down 0.05 (0.01) 0.06 (0.00) 0.744 -1.2 Down 24 .4 4 (2. 27.89 64 lysoPC a C16:0 (2.68) 0.002* 2.5 Up 40.52 (8.99) ) 0.051 1.66 Up 55.91 (16.13) 23.78 (2.09) 0.157 2.35 Up 1. 26 (0. 1.58 13 lysoPC a C16:1 (0.17) 0.071 1.3 Up 1.35 (0.14) ) 0.818 1.07 Up 1.51 (0.17) 1.26 (0.11) 0.219 1.2 Up 2. 00 (0. 3.03 24 lysoPC a C17:0 (0.36) 0.001* 3.11 Up 7.01 (1.97) ) 0.006* 3.51 Up 2.87 (0.94) 1.85 (0.23) 0.533 1.55 Up 19 .4 9 (2. 25.79 33 lysoPC a C18:0 (3.09) 0.001* 2.85 Up 51.51 (11.84) ) 0.024* 2.64 Up 41.15 (12.75) 17.67 (1.70) 0.176 2.33 Up 14 .1 4 (1. 16.09 58 lysoPC a C18:1 (1.54) 0.06 1.29 Up 14.35 (1.50) ) 0.974 1.01 Up 19.33 (3.49) 13.37 (1.38) 0.196 1.45 Up 17 .1 2 (2. 18.49 09 lysoPC a C18:2 (1.87) 0.767 -1.14 Down 10.44 (1.44) ) 0.083 -1.64 Down 12.98 (0.99) 17.43 (2.01) 0.268 -1.34 Down 1. 67 (0. 3.25 17 lysoPC a C20:3 (0.33) 0.668 -1.22 Down 1.92 (0.35) ) 0.717 1.15 Up 0.78 (0.09) 1.82 (0.22) 0.013* -2.33 Down 2. 24 (0. 3.61 18 lysoPC a C20:4 (0.36) 0.071 -1.42 Down 1.92 (0.30) ) 0.447 -1.67 Down 1.18 (0.13) 2.30 (0.26) 0.046* -1.95 Down 0. 61 (0. 0.83 05 lysoPC a C28:0 (0.05) 0.002* -1.93 Down 0.39 (0.10) ) 0.129 -1.56 Down 0.24 (0.05) 0.63 (0.05) 0.001* -2.63 Down 0. 51 (0. 0.75 04 lysoPC a C28:1 (0.10) 0.051 -1.56 Down 0.43 (0.06) ) 0.408 -1.19 Down 0.40 (0.06) 0.53 (0.04) 0.176 1.33 Down 1. 54 (0. 1.86 19 PC aa C28:1 (0.18) 1 1.09 Up 1.20 (0.11) ) 0.447 -1.28 Down 1.99 (0.39) 1.66 (0.15) 0.457 1.2 Up 2. 17 (0. 2.54 26 PC aa C30:0 (0.25) 0.336 1.37 Up 2.44 (0.19) ) 0.243 1.12 Up 3.74 (0.50) 2.28 (0.21) 0.013* 1.64 Up 0. 23 (0. 0.33 04 PC aa C30:2 (0.05) 0.071 2.24 Up 0.59 (0.11) ) 0.010* 2.57 Up 0.59 (0.12) 0.28 (0.05) 0.009* 2.11 Up 7. 06 (0. 8.61 78 PC aa C32:0 (0.75) 0.371 -1.17 Down 5.23 (0.48) ) 0.192 -1.35 Down 6.46 (0.69) 6.68 (0.61) 0.79 -1.03 Down 6. 74 (0. 7.88 81 PC aa C32:1 (0.71) 0.169 -1.36 Down 4.01 (0.19) ) 0.083 -1.68 Down 7.30 (1.09) 6.70 (0.59) 0.656 1.09 Up 5. 99 (0. 8.44 69 PC aa C32:2 (0.89) 0.869 1.1 Up 5.90 (1.16) ) 1 -1.02 Down 5.67 (1.06) 6.33 (0.59) 0.656 -1.12 Down 10 .6 4 (1. 15.78 30 PC aa C32:3 (1.97) 1 1.1 Up 9.74 (2.49) ) 0.767 -1.09 Down 9.03 (1.43) 11.33 (1.12) 0.421 -1.25 Down 99 .9 6 (1 4. 97.65 13 94.61 PC aa C34:1 (8.90) 0.169 -1.26 Down 56.08 (2.82) ) 0.169 -1.78 Down 127.76 (18.00) (10.26) 0.072 1.35 Up 15 4. 36 (2 3. 144.82 31 147.47 PC aa C34:2 (13.13) 0.148 -1.42 Down 60.72 (3.33) ) 0.035* -2.54 Down 154.72 (22.48) (15.38) 0.533 1.05 Up 23 .1 4 (2. 28.61 75 PC aa C34:3 (2.98) 0.083 -1.62 Down 11.89 (1.26) ) 0.035* -1.95 Down 17.28 (2.49) 24.09 (2.44) 0.123 -1.39 Down 3. 86 (0. 7.44 41 PC aa C34:4 (0.91) 0.818 1.02 Up 4.99 (1.28) ) 0.575 1.29 Up 2.66 (0.38) 4.39 (0.55) 0.095 -1.65 Down 4. 40 (0. 6.74 52 PC aa C36:0 (0.77) 0.488 1.16 Up 8.03 (1.70) ) 0.024* 1.83 Up 4.74 (1.03) 4.89 (0.64) 0.976 -1.03 Down 90 .5 0 (1 0. 105.11 75 PC aa C36:1 (9.08) 0.303 -1.12 Down 80.03 (7.23) ) 0.767 -1.13 Down 100.99 (17.81) 87.64 (8.59) 0.656 1.15 Up 15 7. 08 (1 9. 166.65 70 149.78 PC aa C36:2 (14.53) 0.243 -1.22 Down 103.96 (3.55) ) 0.071 -1.51 Down 163.40 (25.32) (14.23) 0.573 1.09 Up 69 .6 8 (8. 79.46 48 PC aa C36:3 (7.61) 0.303 -1.27 Down 45.79 (2.12) ) 0.097 -1.52 Down 70.64 (14.32) 69.02 (6.53) 0.744 1.02 Up 30 .9 6 (3. 39.05 49 PC aa C36:4 (3.45) 0.216 -1.3 Down 21.97 (1.33) ) 0.216 -1.41 Down 34.15 (5.49) 31.91 (2.98) 0.929 1.07 Up 7. 96 (0. 10.24 79 PC aa C36:5 (0.94) 0.112 -1.25 Down 5.80 (0.76) ) 0.243 -1.37 Down 8.17 (1.04) 8.14 (0.82) 0.976 1 Up 2. 13 (0. 3.30 20 PC aa C36:6 (0.33) 0.974 1.08 Up 2.63 (0.61) ) 0.621 1.23 Up 1.85 (0.42) 2.49 (0.24) 0.219 -1.35 Down 1. 28 (0. 1.87 16 PC aa C38:0 (0.20) 0.974 1.01 Up 2.23 (0.48) ) 0.083 1.74 Up 0.91 (0.17) 1.34 (0.17) 0.219 -1.47 Down 2. 84 (0. 4.24 39 PC aa C38:1 (0.62) 0.621 -1.02 Down 5.51 (1.23) ) 0.015* 1.94 Up 2.28 (0.69) 3.22 (0.44) 0.355 -1.41 Down 30 .2 7 (3. 55.71 09 PC aa C38:3 (5.47) 0.974 -1.05 Down 41.21 (3.83) ) 0.129 1.39 Up 23.53 (4.82) 34.70 (4.31) 0.268 -1.47 Down 36 .7 5 (3. 56.15 51 PC aa C38:4 (4.79) 0.668 -1.13 Down 40.97 (2.40) ) 0.767 1.11 Up 34.61 (4.88) 39.73 (4.36) 0.573 -1.15 Down 21 .0 0 (2. 25.35 22 PC aa C38:5 (2.17) 0.575 -1.04 Down 18.51 (1.00) ) 0.519 -1.13 Down 25.72 (3.75) 21.56 (2.14) 0.656 1.19 Up 3. 97 (0. 4.27 52 PC aa C38:6 (0.35) 0.53 -1.04 Down 2.90 (0.21) ) 0.447 -1.37 Down 5.18 (0.86) 4.18 (0.45) 0.457 1.24 Up 0. 20 (0. 0.35 02 PC aa C40:2 (0.05) 0.243 1.23 Up 0.38 (0.07) ) 0.019* 1.9 Up 0.17 (0.02) 0.23 (0.03) 0.387 -1.35 Down 1. 97 (0. 4.41 35 PC aa C40:3 (0.64) 0.53 1.08 Up 4.90 (0.81) ) 0.001* 2.49 Up 0.88 (0.21) 2.17 (0.46) 0.046* -2.47 Down 6. 47 (0. 11.72 84 PC aa C40:4 (1.16) 0.243 1.25 Up 13.93 (1.89) ) 0.004* 2.15 Up 4.47 (0.77) 7.40 (1.13) 0.219 -1.66 Down PC aa C40:5 18.80 0.303 1.22 Up 18.30 (1.69) 14 0.243 1.28 Up 15.40 (2.37) 15.78 (1.65) 0.836 -1.02 Down (1.69) .3 4 (1. 55 ) 3. 17 (0. 3.49 35 PC aa C40:6 (0.27) 0.53 1.12 Up 3.26 (0.17) ) 0.717 1.03 Up 4.31 (0.67) 3.67 (0.40) 0.421 1.17 Up 0. 05 (0. 0.07 00 PC aa C42:1 (0.01) 0.148 1.29 Up 0.09 (0.02) ) 0.019* 1.8 Up 0.05 (0.01) 0.05 (0.00) 0.744 1 Down 0. 08 (0. 0.12 01 PC aa C42:2 (0.01) 0.169 1.25 Up 0.14 (0.01) ) 0.003* 1.75 Up 0.08 (0.01) 0.08 (0.01) 0.692 1 Down 0. 13 (0. 0.25 02 PC aa C42:4 (0.03) 0.717 1 Up 0.27 (0.03) ) 0.002* 2.08 Up 0.06 (0.01) 0.14 (0.03) 0.139 -2.33 Down 0. 54 (0. 0.81 08 PC aa C42:5 (0.09) 0.621 1.11 Up 1.11 (0.27) ) 0.035* 2.06 Up 0.25 (0.06) 0.50 (0.09) 0.196 -2 Down 0. 28 (0. 0.36 03 PC aa C42:6 (0.02) 0.359 1.11 Up 0.44 (0.06) ) 0.007* 1.57 Up 0.26 (0.03) 0.27 (0.02) 0.882 -1.04 Down 0. 56 (0. 0.72 05 PC ae C30:0 (0.06) 0.818 1.06 Up 0.57 (0.05) ) 0.869 1.02 Up 0.65 (0.08) 0.60 (0.06) 0.614 1.08 Up 0. 83 (0. 1.32 10 PC ae C30:1 (0.18) 0.272 1.45 Up 1.59 (0.30) ) 0.006* 1.92 Up 1.56 (0.29) 0.90 (0.11) 0.033* 1.73 Up 3. 10 (0. 3.82 33 PC ae C32:1 (0.37) 0.169 -1.35 Down 2.05 (0.32) ) 0.112 -1.51 Down 2.75 (0.27) 3.07 (0.27) 0.295 -1.12 Down 5. 26 (0. 7.13 68 PC ae C32:2 (0.80) 0.408 -1.25 Down 3.61 (0.57) ) 0.272 -1.46 Down 4.07 (0.80) 5.48 (0.55) 0.196 -1.35 Down 2. 35 (0. 3.66 24 PC ae C34:0 (0.36) 0.974 -1.06 Down 2.85 (0.25) ) 0.303 1.21 Up 1.67 (0.20) 2.49 (0.34) 0.324 -1.49 Down PC ae C34:1 15.63 0.192 -1.24 Down 10.37 (0.41) 11 0.408 -1.14 Down 11.26 (1.68) 11.87 (1.15) 0.614 -1.05 Down (1.42) .8 0 (1. 12 ) 14 .9 2 (1. 18.55 74 PC ae C34:2 (1.95) 0.488 -1.09 Down 11.70 (1.01) ) 0.371 -1.28 Down 15.49 (2.69) 14.90 (1.48) 0.882 1.04 Up 15 .0 6 (2. 19.30 07 PC ae C34:3 (2.50) 0.818 1 Up 10.25 (1.53) ) 0.243 -1.47 Down 15.20 (2.29) 15.57 (1.71) 0.976 -1.02 Down 1. 62 (0. 2.45 18 PC ae C36:0 (0.24) 0.336 -1.29 Down 1.82 (0.15) ) 0.575 1.12 Up 0.92 (0.15) 1.75 (0.20) 0.046* -1.9 Down 12 .8 3 (1. 17.77 18 PC ae C36:1 (1.47) 0.818 -1.02 Down 15.50 (1.45) ) 0.53 1.21 Up 12.04 (2.09) 12.63 (1.23) 0.7 -1.05 Down 17 .1 7 (1. 23.00 59 PC ae C36:2 (2.23) 0.575 -1.1 Down 15.18 (1.39) ) 0.621 -1.13 Down 14.82 (2.63) 16.98 (1.62) 0.494 -1.15 Down 7. 58 (0. 10.60 74 PC ae C36:3 (1.17) 0.53 -1.23 Down 6.46 (0.71) ) 0.447 -1.17 Down 6.20 (0.97) 7.83 (0.75) 0.242 -1.26 Down 6. 13 (0. 9.19 68 PC ae C36:4 (1.09) 0.869 -1.01 Down 5.72 (0.70) ) 0.921 -1.07 Down 5.78 (0.92) 6.37 (0.59) 0.533 -1.1 Down 5. 25 (0. 7.47 49 PC ae C36:5 (0.81) 1 -1.05 Down 4.70 (0.58) ) 0.488 -1.12 Down 5.09 (0.50) 5.56 (0.52) 0.494 -1.09 Down 1. 14 (0. 1.59 09 PC ae C38:0 (0.14) 0.974 1.08 Up 1.57 (0.21) ) 0.129 1.38 Up 1.15 (0.17) 1.27 (0.11) 0.573 -1.1 Down 1. 76 (0. 3.11 23 PC ae C38:1 (0.31) 0.303 1.16 Up 3.31 (0.30) ) 0.006* 1.88 Up 1.60 (0.30) 1.85 (0.28) 0.79 -1.16 Down 2. 31 (0. 3.66 22 PC ae C38:2 (0.35) 0.818 -1.12 Down 2.97 (0.07) ) 0.216 1.29 Up 1.97 (0.53) 2.37 (0.27) 0.421 -1.2 Down PC ae C38:3 6.06 0.575 1.1 Up 5.45 (0.57) 3. 0.010* 1.7 Up 2.49 (0.44) 3.50 (0.49) 0.421 -1.41 Down 21 (0. 33 (0.63) ) 4. 71 (0. 7.48 47 PC ae C38:4 (0.71) 0.974 -1.06 Down 5.73 (0.63) ) 0.303 1.22 Up 3.35 (0.43) 5.06 (0.68) 0.196 -1.51 Down 3. 70 (0. 5.31 34 PC ae C38:5 (0.49) 0.869 -1.08 Down 3.71 (0.30) ) 0.575 1 Up 3.20 (0.43) 4.01 (0.40) 0.242 -1.25 Down 3. 00 (0. 4.02 32 PC ae C38:6 (0.39) 0.818 1.03 Up 2.89 (0.33) ) 0.919 -1.04 Down 3.46 (0.40) 3.47 (0.33) 0.836 1 Down 0. 28 (0. 0.38 03 PC ae C40:1 (0.04) 0.717 1.13 Up 0.37 (0.06) ) 0.216 1.32 Up 0.17 (0.02) 0.29 (0.04) 0.095 -1.71 Down 0. 65 (0. 0.94 06 PC ae C40:2 (0.10) 0.192 1.22 Up 0.93 (0.09) ) 0.083 1.43 Up 0.67 (0.10) 0.71 (0.08) 0.656 -1.06 Down 0. 78 (0. 1.47 08 PC ae C40:3 (0.16) 0.447 1.11 Up 1.25 (0.14) ) 0.015* 1.6 Up 0.56 (0.14) 0.94 (0.15) 0.268 -1.96 Down 1. 18 (0. 2.08 14 PC ae C40:4 (0.20) 0.974 -1.07 Down 1.63 (0.14) ) 0.129 1.38 Up 0.71 (0.13) 1.39 (0.21) 0.139 -1.33 Down 1. 94 (0. 2.71 21 PC ae C40:5 (0.24) 0.53 1.14 Up 2.50 (0.32) ) 0.336 1.29 Up 1.60 (0.24) 2.12 (0.24) 0.268 -1.18 Down 0. 84 (0. 1.05 07 PC ae C40:6 (0.08) 0.818 1.11 Up 0.92 (0.11) ) 0.974 1.1 Up 0.83 (0.12) 0.98 (0.09) 0.355 -1.56 Down 0. 13 (0. 0.20 01 PC ae C42:1 (0.02) 0.243 -1.33 Down 0.11 (0.01) ) 0.169 -1.18 Down 0.09 (0.01) 0.14 (0.02) 0.107 -1.4 Down 0. 14 (0. 0.22 01 PC ae C42:2 (0.02) 0.717 1.09 Up 0.23 (0.03) ) 0.010* 1.64 Up 0.10 (0.02) 0.14 (0.02) 0.242 -1.5 Down PC ae C42:3 0.23 0.408 1.13 Up 0.22 (0.03) 0. 0.019* 1.69 Up 0.10 (0.03) 0.15 (0.02) 0.268 1 Down (0.02) 13 (0. 01 ) 0. 03 (0. 0.05 00 PC ae C44:3 (0.00) 0.336 1.2 Up 0.05 (0.01) 3) 0.035* 1.67 Up 0.03 (0.01) 0.03 (0.00) 0.744 1 Down 0. 06 (0. 0.08 00 PC ae C44:4 (0.004) 0.019* 1.25 Up 0.09 (0.01) 3) 0.007* 1.5 Up 0.05 (0.01) 0.05 (0.01) 0.882 1 Down 0. 05 (0. 0.07 00 PC ae C44:5 (0.01) 0.668 1.14 Up 0.08 (0.01) 4) 0.029* 1.6 Up 0.05 (0.01) 0.05 (0.00) 0.79 1 Down 8. 94 (0. 10.95 90 SM (OH) C14:1 (0.87) 0.097 1.27 Up 8.98 (0.71) ) 0.767 1 Up 10.91 (1.80) 9.73 (0.83) 0.614 1.12 Up 6. 26 (0. 8.48 56 SM (OH) C16:1 (0.81) 0.071 1.35 Up 8.65 (0.83) ) 0.097 1.38 Up 7.03 (1.13) 6.84 (0.65) 1 1.03 Up 11 .9 7 (1. 12.57 76 SM (OH) C22:1 (1.34) 0.051 1.66 Up 12.64 (1.39) ) 0.371 1.06 Up 24.28 (5.19) 12.23 (1.26) 0.016* 1.99 Up 4. 36 (0. 5.55 46 SM (OH) C22:2 (0.55) 0.097 1.41 Up 5.57 (0.52) ) 0.192 1.28 Up 7.29 (1.30) 4.45 (0.41) 0.046* 1.64 Up 1. 00 (0. 1.24 10 SM (OH) C24:1 (0.12) 0.015* 1.72 Up 1.43 (0.13) ) 0.051 1.43 Up 1.50 (0.27) 1.03 (0.09) 0.083 1.46 Up 70 .5 1 (8. 83.03 21 SM C16:0 (7.45) 0.303 1.21 Up 69.02 (4.20) ) 0.717 -1.02 Down 103.53 (15.86) 74.72 (7.00) 0.139 1.39 Up 6. 85 (0. 9.75 70 SM C16:1 (0.86) 0.303 1.2 Up 8.13 (0.42) ) 0.371 1.19 Up 8.66 (1.29) 7.54 (0.69) 0.614 1.15 Up 8. 17 (0. 9.37 87 SM C18:0 (0.85) 0.234 1.3 Up 9.03 (0.65) ) 0.371 1.11 Up 13.05 (2.01) 8.48 (0.81) 0.046* 1.54 Up SM C18:1 3.97 0.488 1.09 Up 3.17 (0.20) 3. 0.575 -1.03 Down 4.36 (0.62) 3.43 (0.34) 0.295 1.27 Up (0.37) 27 (0. 34 ) 0. 17 (0. 0.28 02 SM C20:2 (0.05) 0.013* -4 Down 0.06 (0.03) ) 0.053 -2.83 Down 0.07 (0.03) 0.16 (0.02) 0.041* -2.29 Down 12 .9 4 (1. 17.37 35 SM C24:0 (2.01) 0.112 1.46 Up 21.52 (2.45) ) 0.012* 1.66 Up 19.10 (3.18) 13.85 (1.53) 0.108 1.38 Up 7. 65 (0. 8.08 78 SM C24:1 (0.79) 0.336 1.25 Up 7.04 (1.20) ) 0.668 -1.09 Down 11.29 (2.07) 7.45 (0.80) 0.095 1.52 Up 0. 19 (0. 0.24 02 SM C26:0 (0.02) 0.071 1.63 Up 0.38 (0.05) ) 0.002* 2 Up 0.26 (0.05) 0.19 (0.02) 0.108 1.37 Up 0. 13 (0. 0.10 02 SM C26:1 (0.02) 0.475 2.5 Up 0.14 (0.10) ) 0.262 1.08 Up 0.16 (0.05) 0.12 (0.02) 0.543 1.33 Up 2, 45 8 (2 3,093 02 Hexose (232) 0.019* 1.33 Up 3,268 (219) ) 0.035* 1.33 Up 2,823 (188) 2,354 (179) 0.176 1.2 Up 15 8. 36 (1 0. 195.40 07 135.39 Alanine (10.64) 0.76 1.02 Up 218.02 (18.15) ) 0.032* 1.38 Up 186.68 (31.03) (12.93) 0.235 1.38 Up 10 3. 05 (5. 108.14 91 Arginine (7.43) 0.045* 1.75 Up 162.04 (10.92) ) 0.002* 1.57 Up 133.06 (16.38) 96.03 (7.56) 0.051 1.38 Up 26 .4 5 (2. 30.59 54 Asparagine (3.78) 0.06 -1.87 Down 23.98 (6.38) ) 0.76 -1.1 Down 13.14 (2.88) 26.11 (2.56) 0.012* -1.99 Down 14 .6 0 (3. 16.06 92 Aspartic acid (3.49) 0.144 1.25 Up 14.83 (1.81) ) 0.164 1.02 Up 17.02 (2.17) 12.33 (2.49) 0.083 1.38 Up Citrulline 53.75 0.024* 1.66 Up 86.41 (19.86) 56 0.083 1.53 Up 86.06 (14.79) 51.87 (4.50) 0.011* 1.66 Up (3.47) .3 0 (4. 31 ) 24 7. 88 (2 5. 286.15 27 218.18 Glutamine (21.62) 1 1.06 Up 307.32 (31.07) ) 0.262 1.24 Up 219.57 (12.34) (21.60) 1 1.01 Up 71 .9 0 (7. 90.53 54 Glutamate (7.67) 0.174 1.78 Up 133.03 (31.01) ) 0.072 1.85 Up 105.73 (17.44) 68.42 (6.18) 0.045* 1.54 Up 26 9. 50 (2 6. 208.55 26 252.83 Glycine (16.17) 0.007* 1.55 Up 260.76 (22.69) ) 0.786 -1.03 Down 391.03 (90.27) (34.64) 0.123 1.55 Up 21 1.94 .8 6 (1. 26.94 91 Methionine (3.19) 0.621 1.21 Up 42.43 (15.67) ) 0.659 Up 25.64 (6.43) 19.19 (2.08) 0.171 1.34 Up 30 1.31 .5 5 (2. 40.48 94 Ornithine (2.60) 0.336 -1.37 Down 39.95 (16.22) ) 0.659 Up 27.19 (8.59) 29.23 (3.42) 0.7 -1.08 Down 43 1.61 .7 3 (3. 50.22 87 Phenylalanine (3.84) 0.148 1.29 Up 70.51 (5.21) ) 0.003* Up 43.91 (7.72) 41.71 (3.33) 0.927 1.05 Up 63 1.24 .3 6 (4. 72.70 22 Proline (5.72) 0.668 -1.01 Down 78.25 (3.43) ) 0.062 Up 59.95 (13.67) 57.62 (4.77) 0.7 1.04 Up 66 1.38 .7 6 (6. 66.84 96 Serine (6.58) 0.083 1.62 Up 92.43 (8.45) ) 0.042* Up 85.54 (11.85) 59.38 (5.25) 0.039* 1.44 Up 70 1.16 .6 6 (7. 78.58 01 Threonine (8.16) 0.408 1.04 Up 82.07 (6.01) ) 0.072 Up 74.03 (6.44) 64.22 (6.47) 0.157 1.15 Up Tryptophan 41.67 0.004* -1.97 Down 21.20 (1.55) 29 0.371 -1.39 Down 19.14 (1.66) 29.90 (3.47) 0.095 -1.56 Down (3.33) .4 8 (3. 29 ) 35 1.34 .5 5 (3. 44.12 30 Tyrosine (3.12) 0.974 1 Up 47.71 (5.98) ) 0.097 Up 37.51 (3.32) 35.00 (3.41) 0.79 1.07 Up 18 1.42 5. 31 (1 4. 218.05 62 183.46 Valine (17.60) 0.61 1.11 Up 262.34 (32.90) ) 0.032* Up 206.36 (18.27) (16.27) 0.494 1.12 Up 0. 1.23 60 (0. 0.79 06 Asymmetric dimethylarginine (0.06) 0.415 -1.11 Down 0.74 (0.11) ) 0.272 Up 0.52 (0.06) 0.55 (0.06) 0.465 -1.06 Down 0. 1.5 68 (0. 0.77 06 total Dimethylarginine (0.06) 0.129 1.25 Up 1.02 (0.09) ) 0.024* Up 0.88 (0.05) 0.64 (0.05) 0.023* 1.38 Up 4. 1.77 96 (0. 5.60 33 Carnosine (0.42) 0.032* 1.53 Up 8.76 (1.13) ) 0.004* Up 6.17 (0.95) 4.25 (0.37) 0.107 1.45 Up 68 1.17 .8 8 (3. 77.12 98 Creatinine (6.00) 0.53 1.02 Up 80.77 (6.67) ) 0.185 Up 75.48 (6.12) 57.76 (4.36) 0.063 1.31 Up 3. 1.09 51 (0. 2.74 65 Sarcosine (0.44) 0.019* 1.69 Up 3.81 (0.43) ) 0.336 Up 4.58 (0.72) 2.62 (0.64) 0.016* 1.75 Up 36 1.33 .4 4 (3. 37.70 11 Taurine (2.49) 0.071 1.42 Up 48.38 (6.97) ) 0.072 Up 47.68 (5.44) 33.93 (4.02) 0.095 1.41 Up 1Cows were diagnosed with ketosis (n=6) ranging from wk +1 to +3 (i.e., 5 - 21 d, mean 13 d). 2C16: hexadecanoyl-L-carnitine; C18: octadecenoyl-L-carnitine; C2: acetyl-L-carnitine; C3: propionyl-L-carnitine; C4: butyryl-L-carnitine; C5: valeryl-L-carnitine; lysoPC a: lysophosphatidylcholine acyl; PC aa: phosphatidylcholine diacyl; PC ae: phosphatidylcholine acyl-alkyl; SM (OH): hydroxysphingomyelin; SM: sphingomyelin; lysoPC, PC aa, and PC ae are glycerophospholipids; SM (OH) and SM are sphingolipids. Suppl. Table 4 Concentrations of serum metabolites (mean (SEM)) in healthy control (CON) and ketosis cows at +4, and +8 wks after parturition as determined by DI/LC-MS/MS 8 week after parturition Ketosis / Metabolite1, μM CON P-value F Ketosis / CON Ketosis CON P-value Fold change CON o C10 0.07 (0.03) 0.574 - Down 0.05 (0.01) 0.06 (0.02) 0.378 -1.2 Down 1 C2 1.01 (0.17) 0.015* 1 Up 1.34 (0.02) 1.06 (0.29) 0.394 1.26 Up . C3 0.17 (0.04) 0.818 1 Up 0.21 (0.04) 0.21 (0.07) 0.818 1 Up . C4 0.06 (0.01) 0.297 1 Up 0.07 (0.01) 0.06 (0.01) 0.485 1.17 Up . C5 0.04 (0.01) 0.31 1 Up 0.05 (0.01) 0.05 (0.01) 0.818 1 Up . 36.73 lysoPC a C16:0 20.80 (5.86) 0.004* 5 Up (2.15) 24.87 (6.72) 0.485 1.48 Up . lysoPC a C17:0 1.31 (0.43) 0.009* 4 Up 2.25 (0.14) 2.07 (0.56) 0.818 1.09 Up . 27.33 lysoPC a C18:0 18.63 (6.33) 0.009* 4 Up (2.05) 26.18 (7.37) 0.937 1.04 Up . 26.28 lysoPC a C18:1 13.72 (4.40) 0.015* 2 Up (2.23) 20.72 (5.54) 0.699 1.27 Up . 40.66 lysoPC a C18:2 20.19 (6.89) 0.24 1 Up (3.22) 32.40 (9.30) 0.31 1.25 Up . lysoPC a C20:3 1.48 (0.46) 0.699 - Down 2.30 (0.25) 2.50 (0.65) 0.818 -1.09 Down 1 lysoPC a C20:4 1.37 (0.34) 0.31 1 Up 2.32 (0.18) 2.02 (0.49) 0.589 1.15 Up . lysoPC a C28:0 0.43 (0.06) 0.31 - Down 0.45 (0.05) 0.65 (0.13) 0.18 -1.44 Down 1 lysoPC a C28:1 0.50 (0.16) 0.31 1 Up 0.98 (0.06) 0.86 (0.24) 0.818 1.14 Up . PC aa C28:1 1.61 (0.45) 0.041* 1 Up 4.44 (0.25) 2.99 (0.75) 0.394 1.48 Up . 11.53 PC aa C32:0 4.64 (1.35) 0.041* 1 Up (0.47) 8.12 (2.15) 0.699 1.42 Up . 15.73 PC aa C32:1 5.49 (1.86) 0.093 1 Up (0.46) 9.82 (2.78) 0.24 1.6 Up . 16.62 PC aa C32:2 4.88 (1.73) 0.132 2 Up (2.23) 11.70 (3.31) 0.394 1.42 Up . 36.90 PC aa C32:3 12.97 (4.61) 0.31 1 Up (4.69) 30.98 (9.31) 0.589 1.19 Up . 192.74 PC aa C34:1 73.67 (20.57) 0.026* 2 Up (11.79) 100.50 (31.88) 0.065 1.92 Up . 299.73 PC aa C34:2 148.13 (41.69) 0.18 1 Up (15.05) 240.57 (57.45) 0.818 1.25 Up . 50.23 PC aa C34:3 15.42 (4.81) 0.18 1 Up (3.73) 29.18 (8.14) 0.173 1.72 Up . PC aa C34:4 2.59 (0.89) 0.132 2 Up 9.72 (1.34) 5.78 (1.65) 0.128 1.68 Up 17.62 PC aa C36:0 4.05 (1.44) 0.132 2 Up (2.36) 8.89 (2.80) 0.093 1.98 Up . 172.41 PC aa C36:1 67.97 (22.34) 0.065 1 Up (10.53) 117.92 (31.75) 0.485 1.46 Up . 296.92 PC aa C36:2 150.60 (44.63) 0.31 1 Up (14.41) 269.20 (66.38) 1 1.1 Up . 162.33 PC aa C36:3 60.78 (19.68) 0.24 1 Up (12.30) 118.78 (32.30) 0.575 1.37 Up . 72.33 PC aa C36:4 23.00 (6.74) 0.026* 2 Up (4.35) 44.92 (12.16) 0.18 1.61 Up . 16.06 PC aa C36:5 4.96 (1.39) 0.026* 2 Up (1.45) 10.49 (3.34) 0.485 1.53 Up . PC aa C36:6 1.25 (0.31) 0.026* 2 Up 4.46 (0.59) 3.18 (1.09) 0.485 1.4 Up . PC aa C38:0 1.21 (0.45) 0.31 1 Up 3.31 (0.47) 2.62 (0.81) 0.485 1.26 Up . PC aa C38:1 3.41 (1.77) 0.394 1 Up 7.78 (1.39) 5.63 (1.68) 0.394 1.38 Up . 60.16 PC aa C38:3 23.40 (7.64) 0.18 1 Up (4.86) 54.03 (16.28) 0.937 1.11 Up . 61.26 PC aa C38:4 24.78 (7.10) 0.093 1 Up (3.82) 48.63 (14.52) 0.818 1.26 Up . 41.67 PC aa C38:5 13.40 (3.63) 0.009* 2 Up (4.07) 26.05 (8.37) 0.24 1.6 Up . PC aa C38:6 2.68 (0.75) 0.009* 2 Up 8.61 (0.85) 5.00 (1.54) 0.18 1.72 Up . PC aa C40:2 0.12 (0.04) 0.041* 2 Up 0.53 (0.06) 0.37 (0.12) 0.132 1.43 Up . PC aa C40:3 2.06 (1.02) 0.699 1 Up 6.90 (1.07) 6.06 (2.06) 0.699 1.14 Up . 18.41 PC aa C40:4 5.21 (2.14) 0.041* 1 Up (1.55) 12.38 (3.69) 0.31 1.49 Up . 29.51 PC aa C40:5 9.40 (2.93) 0.015* 2 Up (3.39) 17.57 (5.91) 0.24 1.68 Up . PC aa C40:6 2.52 (0.68) 0.015* 2 Up 6.92 (1.00) 3.67 (1.14) 0.093 1.89 Up . PC aa C42:1 0.03 (0.01) 0.015* 2 Up 0.10 (0.01) 0.06 (0.01) 0.092 1.67 Up PC aa C42:4 0.05 (0.02) 0.394 1 Up 0.19 (0.02) 0.15 (0.05) 0.394 1.27 Up . PC aa C42:5 0.43 (0.21) 0.24 1 Up 1.46 (0.28) 1.02 (0.33) 0.24 1.43 Up . PC aa C42:6 0.31 (0.05) 0.394 1 Up 0.65 (0.08) 0.46 (0.10) 0.18 1.41 Up . PC ae C30:0 0.48 (0.15) 0.041* 1 Up 1.35 (0.09) 0.87 (0.21) 0.31 1.55 Up . PC ae C30:1 1.07 (0.35) 0.093 2 Up 4.23 (0.51) 2.27 (0.75) 0.065 1.86 Up . PC ae C32:1 2.01 (0.67) 0.065 2 Up 5.85 (0.34) 3.85 (0.99) 0.18 1.52 Up . 11.88 PC ae C32:2 4.14 (1.39) 0.24 1 Up (1.09) 8.59 (2.48) 0.485 1.38 Up . PC ae C34:0 1.35 (0.46) 0.093 1 Up 4.07 (0.28) 2.63 (0.71) 0.229 1.55 Up . 43.34 PC ae C34:2 13.70 (4.37) 0.041* 2 Up (2.75) 25.57 (7.02) 0.065 1.69 Up . 49.76 PC ae C34:3 15.82 (5.46) 0.18 1 Up (4.24) 33.61 (9.76) 0.24 1.48 Up . PC ae C36:0 0.85 (0.25) 0.093 1 Up 2.47 (0.30) 1.88 (0.53) 0.589 1.31 Up . 24.22 PC ae C36:1 7.94 (2.37) 0.026* 2 Up (1.65) 17.48 (4.98) 0.589 1.39 Up . 41.92 PC ae C36:2 16.00 (5.22) 0.24 1 Up (3.19) 33.49 (9.83) 0.818 1.25 Up . 16.93 PC ae C36:3 6.55 (2.21) 0.31 1 Up (0.83) 12.71 (3.63) 0.485 1.33 Up . 16.69 PC ae C36:4 5.89 (2.03) 0.31 1 Up (1.18) 9.87 (2.62) 0.041* 1.69 Up . 11.06 PC ae C36:5 3.92 (1.17) 0.041* 1 Up (0.40) 6.72 (2.13) 0.065 1.65 Up . PC ae C38:0 0.90 (0.28) 0.093 2 Up 2.76 (0.36) 1.58 (0.46) 0.31 1.75 Up . PC ae C38:1 0.93 (0.38) 0.093 2 Up 3.10 (0.42) 1.85 (0.533) 0.041* 1.68 Up . PC ae C38:2 2.03 (0.77) 0.394 1 Up 5.32 (0.57) 3.93 (1.16) 0.589 1.35 Up . PC ae C38:3 2.13 (0.71) 0.093 1 Up 6.78 (0.50) 4.75 (1.36) 0.589 1.43 Up . PC ae C38:4 2.40 (0.74) 0.093 1 Up 7.01 (0.30) 4.66 (1.31) 0.24 1.5 Up . PC ae C38:5 2.17 (0.63) 0.041* 2 Up 6.54 (0.26) 4.14 (1.15) 0.065 1.58 Up . PC ae C38:6 2.05 (0.57) 0.015* 2 Up 6.80 (0.53) 4.11 (1.32) 0.132 1.65 Up . PC ae C40:1 0.22 (0.08) 0.18 1 Up 0.51 (0.06) 0.28 (0.08) 0.093 1.82 Up . PC ae C40:2 0.36 (0.11) 0.015* 2 Up 1.32 (0.12) 0.75 (0.20) 0.132 1.76 Up . PC ae C40:3 0.47 (0.17) 0.18 1 Up 1.31 (0.10) 1.00 (0.30) 0.589 1.31 Up . PC ae C40:4 0.63 (0.21) 0.24 1 Up 1.52 (0.12) 1.28 (0.38) 0.589 1.19 Up . PC ae C40:5 1.17 (0.36) 0.065 1 Up 3.42 (0.32) 2.23 (0.69) 0.31 1.53 Up . PC ae C40:6 0.63 (0.19) 0.093 1 Up 1.76 (0.12) 1.10 (0.31) 0.132 1.6 Up . PC ae C42:1 0.07 (0.02) 0.093 1 Up 0.17 (0.01) 0.13 (0.03) 0.699 1.31 Up . PC ae C42:2 0.09 (0.03) 0.132 1 Up 0.25 (0.02) 0.20 (0.07) 0.394 1.25 Up . PC ae C42:3 0.09 (0.03) 0.18 1 Up 0.20 (0.02) 0.16 (0.06) 0.394 1.25 Up . PC ae C44:3 0.03 (0.01) 0.378 1 Up 0.06 (0.01) 0.05 (0.01) 0.394 1.2 Up PC ae C44:5 0.02 (0.004) 0.013* 2 Up 0.07 (0.01) 0.05 (0.01) 0.699 1.4 Up . 20.56 SM (OH) C14:1 7.86 (2.38) 0.065 1 Up (1.25) 11.94 (3.05) 0.065 1.72 Up . 14.90 SM (OH) C16:1 4.68 (1.38) 0.015* 2 Up (1.21) 8.97 (2.80) 0.18 1.66 Up . SM C18:1 2.33 (0.86) 0.015* 2 Up 7.86 (0.54) 5.23 (1.93) 0.31 1.5 Up . SM C20:2 0.10 (0.05) 0.807 1 Up 0.22 (0.06) 0.43 (0.10) 0.18 -1.95 Down . 58.18 SM C24:0 14.80 (4.38) 0.041* 2 Up (4.25) 33.38 (9.40) 0.065 1.74 Up . SM C26:1 0.07 (0.03) 0.065 2 Up 0.07 (0.04) 0.10 (0.04) 0.55 -1.43 Down . Hexose 1,489 (380) 0.026* 1 Up 2,682 (532) 2,228 (633) 0.589 1.2 Up . 186.22 Alanine 120.78 (26.97) 0.041* 1 Up (21.25) 142.52 (31.82) 0.394 1.31 Up . 130.97 Arginine 83.48 (18.62) 0.078 1 Up (15.50) 84.35 (14.63) 0.093 1.55 Up . 25.13 Asparagine 33.66 (6.81) 0.24 - Down (6.02) 37.05 (8.47) 0.24 -1.47 Down 1 17.16 Aspartic acid 38.23 (7.08) 0.132 - Down (2.18) 44.98 (8.71) 0.015* -2.62 Down 1 89.97 Citrulline 55.08 (14.62) 0.132 1 Up (5.31) 66.27 (13.63) 0.589 1.36 Up . 260.78 Glutamine 154.43 (45.07) 0.485 1 Up (45.83) 163.42 (42.98) 0.093 1.6 Up . 122.24 Glutamate 113.15 (32.27) 0.31 1 Up (23.94) 109.50 (27.84) 0.699 1.12 Up . 485.72 Glycine 298.00 (62.05) 0.18 1 Up (38.48) 230.00 (43.46) 0.009* 2.11 Up . 52.32 Histidine 34.99 (8.43) 0.132 1 Up (7.26) 34.70 (6.15) 0.132 1.51 Up . 28.26 Methionine 23.76 (5.84) 0.937 1 Up (7.97) 32.75 (8.75) 0.937 -1.16 Down . 25.69 Ornithine 17.31 (4.34) 0.589 1 Up (7.47) 23.01 (4.61) 0.937 1.12 Up . 105.57 Serine 54.03 (20.71) 0.065 1 Up (8.81) 49.07 (9.53) 0.004* 2.15 Up . 76.58 Threonine 71.38 (18.82) 0.485 1 Up (6.13) 76.85 (19.07) 0.485 1 Down . 19.80 Tryptophan 14.78 (4.54) 0.31 1 Up (1.58) 21.41 (5.33) 1 -1.08 Down . 33.50 Tyrosine 33.26 (11.58) 0.699 1 Up (4.55) 28.79 (7.49) 0.699 1.16 Up . 184.27 Valine 105.88 (21.45) 0.015* 2 Up (25.02) 122.80 (31.22) 0.18 1.5 Up Asymmetric dimethylarginine 0.52 (0.06) 0.688 1 Up 0.46 (0.06) 0.53 (0.02) 0.128 -1.15 Down . total Dimethylarginine 0.72 (0.06) 0.065 1 Up 0.80 (0.04) 0.72 (0.04) 0.24 1.11 Up . 11.10 Carnosine 5.08 (1.05) 0.041* 1 Up (1.75) 7.43 (2.35) 0.173 1.49 Up . 68.19 Creatinine 53.18 (11.38) 0.18 1 Up (3.46) 54.05 (10.98) 0.589 1.26 Up . 56.79 Taurine 26.91 (7.43) 0.093 1 Up (7.51) 24.77 (3.27) 0.002* 2.29 Up . 1C10: decanoyl-L-carnitine; C2: acetyl-L-carnitine; C3: propionyl-L-carnitine; C4: butyryl-L-carnitine; C5: valeryl-L-carnitine; lysoPC a: lysophosphatidylcholine acyl; PC aa: phosphatidylcholine diacyl; PC ae: phosphatidylcholine acyl-alkyl; SM (OH): hydroxysphingomyelin; SM: sphingomyelin; lysoPC, PC aa, and PC ae are glycerophospholipids; SM (OH) and SM are sphingolipids. Suppl. Fig. 1 Suppl. Fig. 2</p>