Figures (5)  Tables (2)

    • Figure 1. 

      Experimental design and pregnancy diagnosis. (a) Experimental design for AI and sample collection for each group. (b) Results for pregnancy diagnosis using rapid visual test kits. (c) The results for pregnancy diagnosis using B mode ultrasound.

    • Figure 2. 

      Analysis of the metabolites detected in the serum samples. (a) PLS-DA analysis score chart. (b), (c) Venn diagrams for sample comparison (P represents the pregnant cows and N represents the non-pregnant cows). (d) Classification of metabolites based on KEGG Compounds analyses. (e) Classification of pathways based on KEGG. (f) Pie chart of compounds classification based on the HMDB database.

    • Figure 3. 

      Analysis of the differential metabolites between pregnant and non-pregnant cows at 28-d post AI. (a) Volcano plot for differential metabolites. The red dots represent the up-regulated metabolites and the blue dots represent the down-regulated metabolites. (b) KEGG classification of the 43 differential metabolites. (c) KEGG pathway enrichment analyses of the 43 differential metabolites. (d) Expression profile of the 43 differential metabolites and their VIP values. (e)−(j) The differential metabolites ranked as the top six based on their VIP values.

    • Figure 4. 

      Proteomic analysis of serum from cows. (a) Venn diagrams for the comparison of samples. (b) Total protein function annotation based on KEGG pathway enrichment analysis. (c) Subcellular localization analysis of the proteins. (d) Volcano plot of proteins detected in cows. The red dots represent the up-regulated proteins and the blue dots represent the down-regulated proteins. (e) Heatmap of all the 18 differential proteins between the two groups. (f) KEGG pathway enrichment analysis for all the 18 differential proteins between the two groups.

    • Figure 5. 

      The level of protein in the serum of pregnant (n = 8), and non-pregnant cows (n = 22) at 28-d post AI examined by ELISA. (a) Level of Apolipoprotein A-Ι (APOA1). (b) Level of Histidine-rich glycoprotein (HRG). (c) Level of Complement factor H (CFH). (d) Level of Myosin heavy chain1 (MYH1). (e) Level of Alpha-2-macroglobulin variant 4 (A2M).

    • Metabolite Regulate p-value FC (P/N)
      Glutamic acid diethyl ester Up 0.009258 1.0222
      2-Amino-3,4-dimethylimidazo[4,5-f]quinoline Down 0.008703 0.7724
      Xi-4-Hydroxy-4-methyl-2-cyclohexen-1-one Down 0.01485 0.983
      4-Heptenoic acid Down 0.03647 0.9874
      CL(i-12:0/i-22:0/i-16:0/i-12:0) Down 0.04905 0.9328
      PC(14:0/18:2(9Z,12Z)) Down 0.02298 0.9549
      Corchorifatty acid F Up 0.0118 1.0192
      12-hydroxyicosanoic acid Down 0.03376 0.9888
      Palmitoylcarnitine Down 0.008085 0.9609
      4-[(2S)-2-Aminopropyl]-N,3-dimethylaniline Down 0.03061 0.9273
      Tetradecanoylcarnitine Down 0.009815 0.916
      Harmalol Down 0.03669 0.9754
      Butyl 4-aminobenzoate Down 0.03791 0.9828
      Cerulenin Up 0.04648 1.0181
      4-(1-Hydroxy-2-methoxyethyl)-5-(hydroxymethyl)-2-methylpyridin-3-ol Down 0.04323 0.9752
      LysoPC(17:0/0:0) Down 0.04554 0.9841
      2-Aminobiphenyl Down 0.03701 0.9748
      3-[[(2S)-2,4-Dihydroxy-3,3-dimethylbutanoyl]amino]propanoic acid Up 0.04388 1.0214
      Caffeic acid 4-O-sulfate Up 0.02259 1.04
      3,4-Dihydro-9-hydroxy-[1]benzothieno[2,3-f]-1,4-thiazepin-5(2H)-one Up 0.04879 1.018
      3-Allylphenol sulfate Up 0.02949 1.0172
      BUTHIDAZOLE Up 0.02255 1.0395
      P-coumaroyltriacetic acid lactone Up 0.01615 1.007
      Pristanal Down 0.04499 0.9827
      Tetracosanedioic acid Down 0.008841 0.9692
      Docosanedioic acid Down 0.04257 0.9799
      2-Hydroxymyristic Acid Down 0.004952 0.9834
      2-hydroxyhexadecanoic acid Down 0.01758 0.9739
      Henagliflozin Up 0.01689 1.1815
      PS (15:0/22:0) Down 0.0001862 0.9553
      PS (15:0/24:1(15Z)) Down 0.00526 0.9687
      Phenprobamate Up 0.04342 1.0463
      2-Naphthalenesulfonic acid Up 0.04935 1.0135
      Esmolol Up 0.02516 1.0356
      2-Carboxy-5,7-dimethyl-4-octanolide Down 0.02919 0.8756
      4-Methyl-1,4-dihydropyridine-3,5-dicarbaldehyde Up 0.03592 1.0178
      Ferulic acid 4-O-sulfate Up 0.03354 1.0312
      3-Methoxy-4-Hydroxyphenylglycol sulfate Up 0.04608 1.0554
      Ascorbic acid 2-sulfate Up 0.04618 1.018
      Lucidone B Up 0.01651 1.0623
      2-Ethylhydracrylic acid Up 0.04302 1.0328
      Pantothenic Acid Up 0.04141 1.0259
      5-Hydroxyindolin-2-one sulfate Down 0.04056 0.9308

      Table 1. 

      Differential metabolites between pregnant and non-pregnant cows at 28-d post AI.

    • Accession number Description Relative abundance (P) Relative abundance (N) FC (P/N) p value (P/N)
      P15497 Apolipoprotein A1 174,509.6198 218,067.4792 0.8003 0.04
      A0A6B9SEB1 Ig lamda chain variable region 15,494.6764 35,129.2435 0.4411 0.03427
      A0A3Q1LUE9 Ig-like domain-containing protein 36,403.3216 7,784.272 4.677 0.000571
      K4JB97 Alpha-2-macroglobulin variant 4 20.7204 23,522.3625 0.000881 2.20E-05
      P19035 Apolipoprotein C-III 3,963.2901 5,955.4715 0.6655 0.01173
      A0A3Q1LS74 Complement factor H 4,146.1947 3,191.1589 1.299 0.01435
      A0A3Q1LU84 Ig-like domain-containing protein 368.7632 838.5748 0.4397 0.008791
      F1MH27 chitinase 452.6277 128.7039 3.517 0.02871
      A0A6B9SE32 Ig lamda chain variable region 271.1981 64.0567 4.234 0.03137
      Q28178 Thrombospondin-1 50.6508 229.5105 0.2207 0.03698
      P20959 Insulin-like growth factor-binding protein 3 86.1332 121.951 0.7063 0.04771
      Q3SYR5 Apolipoprotein C-IV 54.5581 97.5879 0.5591 0.03094
      A0A3Q1LP99 Proteolipid protein 1 12.841 63.9644 0.2008 0.001958
      Q3T0A3 Complement factor D 29.4928 24.0432 1.227 0.04539
      P15467 Ribonuclease 4 31.1259 18.9549 1.642 0.01838
      P15781 Pulmonary surfactant-associated protein B 3.0214 15.637 0.1932 0.001538
      Q9BGU1 Histidine-rich glycoprotein 606.8527 0 2260 0.000058
      A0A3Q1LMB7 Myosin Heavy Chain 1 19.9331 0 2260 0.01072

      Table 2. 

      Differential proteins between pregnant and non-pregnant cows at 28-d post AI.