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Plasma metabolic characterisation of dairy cows with inactive ovaries and oestrus during the peak of lactation

Yu-Xi Song
Yu-Xi Song
, 
Pan Hu
Pan Hu
, 
Yun-Long Bai
Yun-Long Bai
, 
Chang Zhao
Chang Zhao
, 
Cheng Xia
Cheng Xia
 and 
Chuang Xu
Chuang Xu
   | Sep 13, 2019
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Published Online: Sep 13, 2019
Page range: 359 - 367
Received: Feb 01, 2019
Accepted: Jul 16, 2019
DOI: https://doi.org/10.2478/jvetres-2019-0047
Keywords
© 2019 Y.X. Song et al. published by Sciendo
This work is licensed under the Creative Commons Attribution-NonCommercial-NoDerivatives 3.0 License.

Fig. 1

The TIC (total ion count) of the QC (quality control) samples from the IO (inactive ovary cow) and E (oestrous cow) groups by LC–MS; A – ESI+ (positive ion of electrospray ionisation) mode; B – ESI− (negative ion of electrospray ionisation) mode
The TIC (total ion count) of the QC (quality control) samples from the IO (inactive ovary cow) and E (oestrous cow) groups by LC–MS; A – ESI+ (positive ion of electrospray ionisation) mode; B – ESI− (negative ion of electrospray ionisation) mode

Fig. 2

PCA score plot based on LC–MS spectra of plasma of dairy cows obtained from the E (oestrous cow) and IO (inactive ovary cow) groups
PCA score plot based on LC–MS spectra of plasma of dairy cows obtained from the E (oestrous cow) and IO (inactive ovary cow) groups

Fig. 3

The OPLS-DA score plot (a) and permutations test plot (b) based on LC–MS spectra of plasma of dairy cows obtained from the E (oestrous cow) and IO (inactive ovary cow) groups
The OPLS-DA score plot (a) and permutations test plot (b) based on LC–MS spectra of plasma of dairy cows obtained from the E (oestrous cow) and IO (inactive ovary cow) groups

Fig. 4

Pathway analysis of differential plasma metabolites in a – ESI+ (positive ion of electrospray ionisation); and b – ESI− (negative ion of electrospray ionisation) modes between the E (oestrous cow) and IO (inactive ovary cow) groups in LC–MS analysis. The sizes of the bubbles are proportional to the impact of each pathway, with colour denoting the significance from the highest in red to the lowest in white
Pathway analysis of differential plasma metabolites in a – ESI+ (positive ion of electrospray ionisation); and b – ESI− (negative ion of electrospray ionisation) modes between the E (oestrous cow) and IO (inactive ovary cow) groups in LC–MS analysis. The sizes of the bubbles are proportional to the impact of each pathway, with colour denoting the significance from the highest in red to the lowest in white

Fig. 5

Summarised pathways of differential metabolites from the IO (inactive ovary cow) and E (oestrous cow) groups. Down-regulation and up-regulation refer to the IO group compared to the E group. The pathways of differential metabolites were from the Kyoto Encyclopedia of Genes and Genomes database (http://www.kegg.jp) and the MetaboAnalyst 3.0 web server (http://www.metaboanalyst.ca)
Summarised pathways of differential metabolites from the IO (inactive ovary cow) and E (oestrous cow) groups. Down-regulation and up-regulation refer to the IO group compared to the E group. The pathways of differential metabolites were from the Kyoto Encyclopedia of Genes and Genomes database (http://www.kegg.jp) and the MetaboAnalyst 3.0 web server (http://www.metaboanalyst.ca)

Clinical data in two groups of cow samples at 50 to 60 d postpartum for LC–MS detection and for ELISA validation

Parameters Cow samples for LC–MS detection Cow samples for ELISA validation
E IO E IO
No. 16 16 16 16
Age 3.58 ± 1.40 3.07 ± 1.26 3.29 ± 0.34 3.23 ± 0.55
Parity 2.27 ± 1.16 1.88 ± 1.09 1.91 ± 0.28 1.86 ± 0.55
BCS 2.88 ± 0.29 2.69 ± 0.23* 2.84 ± 0.23 2.66 ± 0.21*
MY (Kg/d) 33.56 ± 4.48 34.68 ± 5.00 33.37 ± 4.48 34.90 ± 4.64
Glu (mmol/L) 3.40 ± 0.34 3.36 ± 0.38 3.53 ± 0.51 3.39 ± 0.39
NEFA (mmol/L) 0.21 ± 0.05 0.32 ± 0.12** 0.21 ± 0.08 0.30 ± 0.13*
BHBA (mmol/L) 0.53 ± 0.21 0.63 ± 0.18 0.54 ± 0.17 0.60 ± 0.28

Differential plasma metabolites of IO cows and normal E cows screened by LC–MS

No. Metabolites mzmed rtmed VIP value P value FC Mode
1 Tyramine 120.0801 203.3150 1.0774 0.0078 1.3528 ESI+
2 Betaine 118.0855 288.9015 1.3930 0.0013 1.4872 ESI+
3 L-phenylalanine 166.0855 203.3220 1.0598 0.0099 1.3254 ESI+
4 L-glutamate 148.0594 508.3525 1.7459 0.0059 1.5415 ESI+
5 N6,N6,N6-trimethyl-L-lysine 189.1587 802.7070 1.5786 0.0003 1.4007 ESI+
6 D-proline 116.0703 315.4230 1.4542 0.0007 1.2543 ESI+
7 L-pyroglutamic acid 130.0490 508.3120 1.4025 0.0050 1.2130 ESI+
8 Cholic acid 391.2832 46.9290 1.8280 0.0003 0.6327 ESI+
9 p-chlorophenylalanine 200.0463 243.4400 1.2859 0.0024 0.7587 ESI+
10 L-phenylalanine 164.0719 203.9900 1.2468 0.0144 1.4683 ESI−
11 L-citrulline 349.1815 485.9170 1.1153 0.0177 1.6022 ESI−
12 L-glutamate 146.0457 510.6010 1.8904 0.0085 1.4992 ESI−
13 γ-L-glutamyl-L-valine 245.1133 468.3435 1.6628 0.0038 1.5038 ESI−
14 Arachidic acid 311.2936 50.1280 1.5495 0.0431 0.6241 ESI−
15 L-leucine 130.0878 219.6660 1.4373 0.0073 1.4759 ESI−
16 L-valine 116.0717 290.1260 1.5806 0.0032 1.5261 ESI−
17 L-pyroglutamic acid 128.0356 281.0020 1.9003 0.0025 1.6062 ESI−
18 L-isoleucine 130.0877 240.2465 1.7346 0.0010 1.6988 ESI−
19 L-alanine 88.0408 388.2320 1.9019 0.0000 1.3377 ESI−
20 D-proline 114.0560 315.8370 1.9989 0.0003 1.3718 ESI−
21 Cytidine 302.0975 208.5070 1.1421 0.0218 1.3280 ESI−
22 Creatine 261.1303 388.7810 1.5459 0.0008 1.6097 ESI−
23 Phenylacetylglycine 192.0663 103.3500 1.7514 0.0001 1.7482 ESI−
24 α-N-phenylacetyl-L-glutamine 385.1378 103.4960 1.6689 0.0003 1.8358 ESI−

The ELISA validation of 5 differential metabolites between normal oestrous cows and inactive ovary cows

Parameters E IO
No. 16 16
Cholic acid, pmol/L 59.85 ± 11.56 52.75 ± 11.88
D-proline, ng/mL 37.84 ± 5.06 41.53 ± 5.14*
L-alanine, ng/mL 91.44 ± 7.24 97.52 ± 7.12*
L-glutamate, ng/mL 22.50 ± 3.60 24.07 ± 2.72
L-pyroglutamic acid, ng/mL 3.23 ± 0.52 3.60 ± 0.48*
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