The present study was aimed to establish a novel TaqMan real-time PCR (RTm-PCR) for detecting and typing bovine viral diarrhea virus (BVDV), and also to develop a diagnostic proto- col which simplifies sample collection and processing. Universal primers and TaqMan-MGB probes were designed from the known sequences of conserved 5′ - and 3′-untranslated regions (5’UTR, 3’UTR) of the NADL strain of BVDV. Prior to optimizing the assay, cDNAs were tran- scribed in vitro to make standard curves. The sensitivity, specificity and stability (reproducibility) were evaluated. The RTm-PCR was tested on the 312 feces specimens collected from persistently infected (PI) calves. The results showed the optimum conditions for RTm-PCR were 17.0 μmol/L primer, 7.5 μmol/L probe and 51.4°C annealing temperature. The established TaqMan RTm-PCR assay could specially detect BVDV without detecting any other viruses. Its detection limit was 1.55×100 copies/μL for viral RNA. It was 10000-fold higher than conventional PCR with excel- lent specificity and reproducibility. 312 samples were tested using this method and universal PCR from six dairy farms, respectively. Positive detections were found in 49 and 44 feces samples, respectively. The occurrence rate was 89.80%. In conclusion, the established TaqMan RTm-PCR could rapidly detect BVDV and effectively identify PI cattle. The detection limit of RTm-PCR was 1.55 copies/μL. It will be beneficial for enhancing diagnosis and therapy efficacy and reduce losses in cattle farms.
The drainage consolidation method has been efficiently used to deal with soft ground improvement. Nowadays, it has been suggested to use a new sand soil which is a composite of sand and recycled glass waste. The permeability performance of glass-sand soil was explored to judge the feasibility of glass-sand soil backfilled in the drainage consolidation of sand-drained ground. For comparison purposes, different mix proportions of recycled glass waste, fineness modulus, and glass particle size were analyzed to certify the impact on the permeability coefficient and the degree of consolidation. The numerical results show that adding a proper amount of recycled glass waste could promote the permeability performance of glass-sand soil, and the glasssand soil drain could be consolidated more quickly than a sand drain. Experiments showed that glass-sand soil with the a 20% mix of recycled glass waste reveals the optimum performance of permeability.