Establishment and application of a PCR assay for the identification of virulent and attenuated duck plague virus DNA in cotton swabs.

Duck plague is an acute, febrile, and septic infectious disease caused by duck plague virus (DPV), which causes serious harm to the duck industry in China. Ducks latently infected with DPV display a clinically healthy state, which is one of the epidemiological characteristics of duck plague. In the present study, to rapidly distinguish vaccine-immunized ducks from wild virus-infected ducks during production, a PCR assay based on the newly identified LORF5 fragment was developed to effectively and accurately identify viral DNA in cotton swab samples and was used to assess artificial infection models and clinical samples. The results showed that the established PCR method had good specificity and that only the virulent and attenuated DNA of duck plague virus was specifically amplified, as the results for the detection of common duck pathogens (duck hepatitis B virus, duck Tembusu virus, duck hepatitis A virus type 1, novel duck reovirus, Riemerella anatipestifer, Pasteurella multocida, and Salmonella) were negative. The amplified fragments of virulent and attenuated strains were 2,454 bp and 525 bp, and their minimum detection amounts were 0.46 pg and 46 pg, respectively. The detection rate of the virulent and attenuated DPV strains in duck oral and cloacal swabs was lower than that of the gold standard PCR method (GB-PCR, which is unable to distinguish virulent and attenuated strains), and cloacal swabs from clinically healthy ducks were more suitable for detection than oral swabs. In conclusion, the PCR assay established in the present study can be used as a simple and effective method for the clinical screening of ducks that are latently infected with virulent strains of DPV and shedding virus, which can provide technical support for the elimination of duck plague from duck farms.

Adaptive Neural Network-Based Event-Triggered SOC Observer With Application to a Stochastic Battery Model.

Accurate state of charge (SOC) is crucial to achieving safe, reliable, and efficient use of batteries. This article proposes an adaptive neural network (NN)-based event-triggered observer to estimate SOC. First, a stochastic battery equivalent circuit model (ECM) is established, where an adaptive NN is employed to approximate the unknown nonlinear part. The learning process of network weight is conducted online to observe the variations of model parameters and avoid time-consuming processes for parameter extraction. Besides, for the purpose of saving computational cost, an event-triggered mechanism (ETM) is employed in the weight updating law, which means the weights only update when it is necessary. Then, an adaptive radial basis function (RBF) NN-based SOC observer is designed, and its stability is proven by the Lyapunov theory. Moreover, the strictly positive lower bound of interevent time is derived, and undesirable Zeno behavior can be excluded. Finally, the accuracy and robustness of the proposed observer are evaluated by experiments and simulations. Results show that the proposed method can estimate SOC accurately in the presence of initial deviation and sensor noises.

Advance of Clustered Regularly Interspaced Short Palindromic Repeats-Cas9 System and Its Application in Crop Improvement.

Clustered regularly interspaced short palindromic repeats (CRISPR)-Cas9 is the third generation of novel targeted genome editing technology after zinc finger nucleases (ZFNs) and transcription activator like effector nucleases (TALENs). It is also one of the most promising techniques for mutating and modifying genes. The CRISPR-Cas9 system has the advantages of simplicity, high efficiency, high specificity, and low production cost, thus greatly promoting the study of gene function. Meanwhile, it has attracted the attention of biologists. After the development and improvement in recent years, CRISPR-Cas9 system has become increasingly mature and has been widely used in crop improvement. Firstly, this review systematically summarizes the generation and advantages of CRISPR-Cas9 system. Secondly, three derivative technologies of the CRISPR-Cas9 system are introduced. Thirdly, this review focuses on the application of CRISPR-Cas9 system in gene knockout, gene knock-in, and gene regulation, as well as the improvement of yield, quality, and biological resistance of important crops such as rice, wheat, soybean, corn, and potato. Finally, this review proposes the potential challenges of CRISPR-Cas9 system, and discusses the future development of CRISPR-Cas9 system.