Application of proline in prevention and treatment of honeybee virus infection
A technology of virus infection and proline, applied in the field of bee breeding, can solve the problems of low production efficiency, limitation of mass production of antimicrobial peptides, high production cost, etc., and achieve the effect of reducing copy number, helping normal growth and development, and inhibiting proliferation
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Embodiment 1
[0043] Example 1 Intervention of healthy Chinese honeybee larvae and infected larvae with proline
[0044] 1. For sample book
[0045] The Chinese honey bee comes from the experimental colony of the Institute of Bee Research, Chinese Academy of Agricultural Sciences. In order to obtain 2-day-old larvae, the queen bee was kept on the spleen and laid eggs for 96 hours. The spleen containing the 2-day-old larvae was taken out from the colony and placed in a 24-well plate, and placed at a temperature of 32±1℃ and relative humidity. Pre-cultivation is performed in a constant temperature and humidity incubator of 75±5% RH.
[0046] The reverse transcription-polymerase chain reaction (RT-PCR) method was used to detect the presence of CSBV and other viruses in the test larvae (including black queen bee virus BQCV, inferior virus DWV, Israeli paralysis virus IAPV, Kashmir virus KBV, acute paralysis virus ABPV, Chronic paralysis virus (CBPV) infection, and healthy larvae without virus infect...
experiment example 1
[0068] Experimental example 1 Analysis of survival rate and mortality of healthy larvae and infected larvae with proline intervention
[0069] 1. Survival rate
[0070] The survival rate statistics of each group of bee larvae in Example 1 are as follows: figure 1 As shown, the results show that the survival rate of the larvae in the conventionally fed CK group and the preparation group can reach more than 86%, indicating that the proline intervention will not cause abnormal death of healthy larvae, and will not affect the growth and development of healthy larvae.
[0071] A large number of larvae in the infected group died, and the survival rate was only 43.05%. The survival rate of larvae in the proline intervention group was greatly increased to more than 77%. Among them, the high concentration group (75μg / mL) had the highest survival rate of 6-day-old larvae, reaching 88.89%, the results show that the use of proline intervention can prevent about 50% of the infected larvae from di...
experiment example 2
[0079] Experimental example 2 Proline intervention in healthy larvae and virus copy number detection after infected larvae
[0080] The apparently healthy 4-6-day-old mid-bee larvae (CK group) in Example 1 were tested for CSBV, and it was found that the virus copy number of each day-old larva was 66-70, indicating that the apparently healthy larvae also had virus infection, but No obvious symptoms are shown, indicating that the virus is generally invisible in the bee colony. The larvae can live with the virus within a certain concentration. This is similar to Liu Shan (Shan,L.,Liuhao,W.,Jun,G.,Yujie). ,T.,Yanping,C.,Jie,W.,Jilian,L.,ChineseSacbrood Virus Infection in Asian Honey Bees(Apis cerana cerana) and HostImmune Responses to the Virus Infection,Journal of InvertebratePathology.2017, doi: http: / / dx.doi.org / 10.1016 / j.jip.2017.09.006) results are consistent. After proline intervention, the virus copy number in the apparently healthy 4--6 day-old mid-bee larvae (preparation g...
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