Identification method of laying duck stress state
A technology of stress state and laying ducks, which is applied in the field of identification of stress state of laying ducks, can solve the problems of complex sampling, difficult measurement, and inability to realize live evaluation, etc., achieves the effect of simple analysis and improved practical significance
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Embodiment 1
[0022] Example 1 is used to identify the acquisition of marker metabolites of the stress state of laying ducks
[0023] 1. Take 500 μl of blood from laying ducks with known cage days as the experimental group (group to be identified), and take 500 μl of blood from laying ducks that are similar in age to caged ducks (±8 days) as the control group. Blood samples were anticoagulated with sodium heparin, centrifuged at 5000 rpm, 4°C for 10 min, and 200 μL of plasma was obtained, which was frozen in liquid nitrogen and stored for later use.
[0024] 2. Take part of the plasma samples from step 1 and mix them in equal amounts to prepare a quality control sample. Put the plasma sample obtained in step 1 and the quality control sample at 100 μL / sample into an EP tube, add 500 μL of 80% methanol aqueous solution containing 0.1% formic acid, After vortexing, let stand in an ice bath for 5 minutes, centrifuge at 15,000 rpm and 4°C for 10 minutes, take a certain amount of supernatant and ...
Embodiment 2
[0033] 1. Identification of stress state of Jingjiang laying ducks
[0034] Step 1. Obtaining of samples: select 120-day-old Jingjiang laying ducks of the third line of supporting commercial egg ducks, and collect blood samples of laying ducks in the 5-day group, 10-day group, 15-day group and the ground level raising group. Group of 6 samples. Blood samples were anticoagulated with sodium heparin, centrifuged at 5000 rpm, 4°C for 10 min, and 200 μL of plasma was obtained, which was frozen in liquid nitrogen and stored for later use.
[0035] Step 2. Chromatography-mass spectrometry analysis: Take part of the plasma samples in step 1 and mix them in equal amounts to prepare quality control samples. Put 100 μL / sample of the plasma samples and quality control samples obtained in step 1 into EP tubes, add 500 μL containing 0.1% formic acid 80% methanol aqueous solution, after vortex shaking, let stand in ice bath for 5min, centrifuge at 15000rpm, 4°C for 10min, take a certain am...
Embodiment 3
[0049] 1. Identification of stress state in mountain shelducks
[0050] Step 1. The species is mountain shelduck, the age of laying ducks in baskets is 100 days, and the evaluation period is the 6th day and the 12th day after basketing. Collect the laying duck samples to be evaluated on the 6th day and the 12th day after being caged, and at the same time collect 6 samples from each group of laying duck samples in the ground culture group of similar age. Blood samples were anticoagulated with sodium heparin, centrifuged at 5000 rpm, 4°C for 10 min, and 200 μL of plasma was obtained, which was frozen in liquid nitrogen and stored for later use.
[0051] Step 2. Take 100 μL / sample of the obtained plasma sample and quality control sample, put it in an EP tube, add 500 μL of 80% methanol aqueous solution containing 0.1% formic acid, vortex, let stand in an ice bath for 5 minutes, and centrifuge at 15,000 rpm and 4°C 10min, take a certain amount of supernatant and dilute it with ma...
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