Wickerhamomyces anomalus for feeding and application thereof
A Wickham and abnormal technology, applied in the field of abnormal Wickham yeast for feed, can solve the problem of less research and application of Wickham yeast
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Embodiment 1
[0027] Example 1 Isolation and identification of abnormal Wickham yeast (Wickerhamomyces anomalus) Y4
[0028] 1. Identification of strain Y4
[0029] 1. Isolation and cultivation of strains
[0030] Take 1g of the soil sample near the pig breeding base of the West Campus of China Agricultural University in Haidian District, Beijing, and put it into a test tube with 9ml of normal saline, vortex and shake it to make a 1:10 dilution, and then take the dilution to increase by 10 times Dilute, and then select 1ml of each of 3 appropriate gradient dilutions and spread them on the YPD medium. Cultivate at 30°C for 48-72 hours, observe and record the colony shape, pick a single colony with good growth, and separate and purify by streaking.
[0031] 2. Methylene blue staining of strains
[0032] Drop a drop of sterilized distilled water on the slide, and pick a single colony (see figure 1 ) in water, add 1 drop of methylene blue dye solution, and cover with a cover glass; observe ...
Embodiment 2
[0043] Example 2 Stress resistance detection of abnormal Wickham yeast Y4
[0044] 1. Heat resistance test: Put the abnormal Wickham yeast Y4 (CGMCC No.16540) bacterial liquid in the water bath for 20 minutes, and treat it at 60°C, 80°C, and 100°C respectively, and repeat each treatment 3 times. After the end, the number of viable bacteria was determined by pouring method.
[0045] The survival rate of Wickhamiana anomaly Y4 CGMCC No.16540 can reach 99% after treatment at 60°C for 20 minutes, over 95% after treatment at 80°C for 20 minutes, and over 90% after treatment at 100°C for 20 minutes .
[0046] 2. Acid resistance test: Inoculate Wickhamiana anomaly Y4 into YPD medium with pH values of 2.0, 3.0, and 4.0, respectively, and measure the number of viable bacteria by plate pouring method at 1h, 2h, 3h, and 4h respectively.
[0047] Abnormal Wickham yeast Y4 CGMCC No.16540 can grow normally in the environment of pH 4, the results are shown in Figure 4 . When the pH wa...
Embodiment 3
[0056] Example 3 Determination of growth curve of abnormal Wickham yeast Y4
[0057] The growth curve represents the dynamic change of the strain in the whole process of growth and reproduction in a new suitable environment until aging and death. Abnormal Wickham yeast Y4 CGMCC No.16540 was inoculated into YPD medium at an inoculation amount of 10% (v / v), and cultured at 30° C. for 46 hours. The YPD medium without bacterial liquid was used as a blank control. Measure the OD600 value every 2-6 hours to calculate the number of viable bacteria. The experiment was repeated three times, the results were averaged, the data were recorded and the growth curve was drawn. Such as Figure 6 As shown, at 0-18 hours, Wickhamiana anomaly Y4 was in the logarithmic growth phase with a high reproduction rate. Abnormal Wickhamiana Y4 numbers stabilized at 18-46 hours. After 46 hours, the growth rate decline phase was entered.
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