Study On Novel Rapid Diagnostic Methods For Bovine Diseases And Feed Safety

There are many challenges for the development of modern animal husbandry,and the key is setting up the Animal Husbandly Safety System,including Bio-security System,Animal-based Food Safety System and Ecological Environment Safety.The core of Bio-security System is the prevention and control for animal disease,and the suitable diagnostic kits are required as the base of other work for the Bio-security System,the epidemiological survey data,epidemic trend of diseases,vaccination performance evaluation can be shown with the detection results with the kits,even the diagnostics of disease and source finding of disease can be done with test results.The source of the food safety is feed safety management and veterinary drug management,and the diagnostic kits that are fit for the feed safety and veterinary drug testing,and the test result is the base of management.With the Bio-security Safety system,feed safety management and veterinary drug management,the Ecological Environment Safety System will be optimized.Mycoplasma Bovis is a widely spreaded disease in China and many other countries,and M.Bovis is a microorganism that will cause mammitis,arthritis,pneumonia,conjunctivitis in the cattle,and abortion in pregnant cattle,and the animal farming efficiency will be decreased.Bovine Viral Diarrhea Virus(BVDV)is classified into genus Pestivirus within the Flaviviridae family.The typical symptoms of the B VDV infection are diarrhoea,fever,followed by reduction in the milk production.The immune suppressive effect of BVDV may potentiate infection by other microorganisms.Because of the existence of persistently infectious(PI)animals,BVDV is not easy to eliminated.Clenbuterol is forbidden to add into feed,and human will intake it from the animal-based food,then human health will be damaged.Aflatoxin B1(AFB1)is a deadly carinogenic material,and always generated by Aspergillus flavus in the plants,grains and feed.The AFB1 in feed will be taken by human,and harm the human health.This study is to develop serial point of care diagnostic kits for above 2 pathogens,and 2 harmful substances,employing Lateral Flow technology,Colloid Gold technology and Up-converting Phosphor Technology(UPT).UPT is a new technology,to detect target quantitatively with the up-converting phosphor nano-beads,which are composed with rare earth elements.UPT has the advantages,to develop the diagnostic kits with low-background,quantitative test results and stability.With UPT technology,the test value is related with the concentration of the target objective,then the concentration can be calculated with the test value.Finally we developed the M.bovis diagnostic kit(P48-CG),BVDV POC kit(BVDV-CG),clenbuterol UPT kits(CL-UPT)and AFB1 UPT kits(AFB1-UPT).The accordant rate of P48-CG and commercial ELISA kit is above 93%,and the sensitivity is the same with ELISA kit.619 serum/plasma samples were tested with BVDV-CG,sensitivity and specificity is 98.4%and 99.4%respectively;for 858 whole blood samples,96.5%and 99.1%;for 440 ear notch samples,100%and 100%.The sensitivity of CL-UPT is 0.05ppb,and the accordant rate with LC-MS/MS is above 85%.The sensitivity of AFB1-UPT is 0.39ppb,and the accordant rate with HPLC is above 85%.The serial diagnostic kits,for animal disease prevention and control,feed safety,will be the tools to optimize the animal husbandry safety system.