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Abstract

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.
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Abstract

The Intrauterine fetal development process is complicated and affected by many regulating factors such as maternal nutritional status, transcription factors and adipokines. Adipokines are kinds of active substances secreted by adipose tissue, including more than 50 kinds of molecules. To explore the correlation between calf birth weights and adipokines including adiponectin, leptin, visfatin, and IGF-1 in cows venous and venous cord blood. Fifty-four healthy multiparous Chinese Holstein cows were used; in which, cows with a calf weight less than 40 kg were included in group A (n=9); those with a calf weight between 40 kg~45 kg were included in group B (n=25) and ≥45 kg were included in group C (n=20), venous blood and cord venous blood was collected. An ELISA kit was used to evaluate the concentration of adiponectin, leptin, visfatin, and IGF-1, correlations between index-index and index-calf birth weight were analysed. In both cows venous and cord venous blood, adiponectin, leptin, visfatin, and IGF-1 levels were significantly correlated with each other (p<0.01), and levels of these adipokines in venous blood were significantly higher than cord venous blood (p<0.01). Adiponectin, leptin, visfatin, and IGF-1 in venous cord blood were positively correlated with calf birth weights, and significantly correlated with calf birth weights respectively (p<0.01). Our study showed that adiponectin, leptin, and IGF-1 were found in venous blood and cord venous blood, and adiponectin, leptin, and IGF-1 in venous and cord venous blood potentially inter-regulated each other; adiponectin, leptin, and IGF-1 in venous blood were not significantly correlated with calf birth weights, while adiponectin, leptin, visfatin, and IGF-1 in venous cord blood were significantly correlated with calf birth weights, respectively.
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