“…The normal control oestrus (CON) group (n = 50): Cows with a low plasma BHBA content (>1.20 mmol/L) at 14–21 days postpartum were in a positive energy balance state, tracked to 60 days postpartum and had spontaneously oestrous signs 13,14 . The results of the rectal examination, B-ultrasonography, and follicular diameter measurements of the oestrous cows were the opposite of those in the cows with inactive ovaries 13,15 . The oestrous cows were assigned to the CON group, and then, the background data collected for the IO group were also collected for the CON group.…”
Section: Methodsmentioning
confidence: 64%
“…The experimental animal selection and grouping are shown in Fig. 1 according to the criteria of postpartum inactive ovaries and oestrus in dairy cows 13–15 .Figure 1Schematic of the selection of experimental animals.…”
Section: Methodsmentioning
confidence: 99%
“…The inactive ovary (IO) group (n = 50): Cows with a high plasma BHBA content (>1.20 mmol/L) at 14–21 days postpartum were in a negative energy balance state, were tracked to 60 days postpartum and had no oestrous signs 13,14 . A rectal examination and B-ultrasound examination were performed at 60–90 days postpartum by a trained professional; if the cow had no follicular development or follicular retardation (less than 4 mm) for 7 days 13,15 , it was assigned to the IO group, and then, all background data, including age, parity, milk yield, disease, and body condition, were collected. If these cows underwent oestrus between 60 and 90 days postpartum and experienced other diseases after calving, they were removed from the IO group.…”
Section: Methodsmentioning
confidence: 99%
“…Five plasma samples from each group were mixed to become a mixed sample, to which lysis buffer was added. The mixed plasma sample was placed under sonication for 60 seconds with a frequency of 0.2 seconds and an amplitude of 22%, extracted at room temperature for 30 minutes, and centrifuged at 20 °C for 20 minutes at 15,000 × g. The supernatant was then carefully collected to determine its protein concentration by the Bradford method 15 . In this experiment, a human plasma/plasma high abundance protein kit (MERCK, USA) was used to remove high abundance proteins in the mixed supernatant samples for iTRAQ-labelling proteomics experiments.…”
Section: Methodsmentioning
confidence: 99%
“…The supernatant was then drawn to load 10 μL of the supernatant sample. The loading pump flow rate was 350 nL/min, for 15 minutes, and the separation flow rate was 350 nL/min 15 .…”
To screen differentially expressed proteins in the blood dairy cows with inactive ovaries caused by a negative energy balance and to determine the roles of the identified proteins in the development of inactive ovaries.Holstein cows at 14 to 21 days postpartum in an intensive dairy farm were examined for their energy balance (EB) status by blood β-hydroxybutyrate (BHBA) and assigned to the inactive ovary (IO) group (n = 50) and the normal oestrus control (CON) group (n = 50) at 60 to 90 days postpartum by means of the oestrus manifestation, rectal examination and B-ultrasound examination. Fourteen differentially expressed proteins from 61 proteins in the plasma of dairy cows with IOs were identified by iTRAQ/LC-MS/MS and GO, KEGG, and PATHWAY analysis. Eleven expressed proteins were upregulated, and 3 expressed proteins were downregulated. Among the 10 differentially expressed proteins verified by Western blot or ELISA, the relative expression levels of ALDOB, IGFBP2, ITIH3 and LDHB in mixed samples and single samples were consistent with the proteomic protein results. PKM2, GPX3, ALDOB, RBP4 and AHSG were significantly different between the two groups (P < 0.05); APOA4 and SPAM1 were not significantly different (P > 0.05) but were still downregulated in the ovarian resting group. This study confirmed that 14 plasma differential proteins in the inactive ovaries of postpartum dairy cows were associated with follicular development, and these findings provide a foundation for further research on the mechanism and prevention of inactive ovaries in dairy cows.
“…The normal control oestrus (CON) group (n = 50): Cows with a low plasma BHBA content (>1.20 mmol/L) at 14–21 days postpartum were in a positive energy balance state, tracked to 60 days postpartum and had spontaneously oestrous signs 13,14 . The results of the rectal examination, B-ultrasonography, and follicular diameter measurements of the oestrous cows were the opposite of those in the cows with inactive ovaries 13,15 . The oestrous cows were assigned to the CON group, and then, the background data collected for the IO group were also collected for the CON group.…”
Section: Methodsmentioning
confidence: 64%
“…The experimental animal selection and grouping are shown in Fig. 1 according to the criteria of postpartum inactive ovaries and oestrus in dairy cows 13–15 .Figure 1Schematic of the selection of experimental animals.…”
Section: Methodsmentioning
confidence: 99%
“…The inactive ovary (IO) group (n = 50): Cows with a high plasma BHBA content (>1.20 mmol/L) at 14–21 days postpartum were in a negative energy balance state, were tracked to 60 days postpartum and had no oestrous signs 13,14 . A rectal examination and B-ultrasound examination were performed at 60–90 days postpartum by a trained professional; if the cow had no follicular development or follicular retardation (less than 4 mm) for 7 days 13,15 , it was assigned to the IO group, and then, all background data, including age, parity, milk yield, disease, and body condition, were collected. If these cows underwent oestrus between 60 and 90 days postpartum and experienced other diseases after calving, they were removed from the IO group.…”
Section: Methodsmentioning
confidence: 99%
“…Five plasma samples from each group were mixed to become a mixed sample, to which lysis buffer was added. The mixed plasma sample was placed under sonication for 60 seconds with a frequency of 0.2 seconds and an amplitude of 22%, extracted at room temperature for 30 minutes, and centrifuged at 20 °C for 20 minutes at 15,000 × g. The supernatant was then carefully collected to determine its protein concentration by the Bradford method 15 . In this experiment, a human plasma/plasma high abundance protein kit (MERCK, USA) was used to remove high abundance proteins in the mixed supernatant samples for iTRAQ-labelling proteomics experiments.…”
Section: Methodsmentioning
confidence: 99%
“…The supernatant was then drawn to load 10 μL of the supernatant sample. The loading pump flow rate was 350 nL/min, for 15 minutes, and the separation flow rate was 350 nL/min 15 .…”
To screen differentially expressed proteins in the blood dairy cows with inactive ovaries caused by a negative energy balance and to determine the roles of the identified proteins in the development of inactive ovaries.Holstein cows at 14 to 21 days postpartum in an intensive dairy farm were examined for their energy balance (EB) status by blood β-hydroxybutyrate (BHBA) and assigned to the inactive ovary (IO) group (n = 50) and the normal oestrus control (CON) group (n = 50) at 60 to 90 days postpartum by means of the oestrus manifestation, rectal examination and B-ultrasound examination. Fourteen differentially expressed proteins from 61 proteins in the plasma of dairy cows with IOs were identified by iTRAQ/LC-MS/MS and GO, KEGG, and PATHWAY analysis. Eleven expressed proteins were upregulated, and 3 expressed proteins were downregulated. Among the 10 differentially expressed proteins verified by Western blot or ELISA, the relative expression levels of ALDOB, IGFBP2, ITIH3 and LDHB in mixed samples and single samples were consistent with the proteomic protein results. PKM2, GPX3, ALDOB, RBP4 and AHSG were significantly different between the two groups (P < 0.05); APOA4 and SPAM1 were not significantly different (P > 0.05) but were still downregulated in the ovarian resting group. This study confirmed that 14 plasma differential proteins in the inactive ovaries of postpartum dairy cows were associated with follicular development, and these findings provide a foundation for further research on the mechanism and prevention of inactive ovaries in dairy cows.
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