Epidemiological and molecular analysis of avian influenza A(H7N9) virus in Shanghai, China, 2013-2017.

BACKGROUND: Human infections with a novel avian influenza A virus (H7N9) were reported in Shanghai municipality, China, at the beginning of 2013. High-pathogenic avian influenza (HPAI) H7N9 virus emerged in late February 2017 along with existing low-pathogenic avian influenza (LPAI) H7N9 virus, and this has the potential to develop into a pandemic that could be harmful to humans. METHODS: To elucidate the epidemiological characteristics of H7N9-infected cases from 2013 to 2017 in Shanghai, data on the 59 laboratory-confirmed human cases and 26 bird and environmental contamination cases were collected from the WHO website and Food and Agriculture Organization Emergency Prevention System for Animal Health (FAO EMPRES-AH). Full-length sequences of H7N9 viruses that emerged in Shanghai were collected from the Global Initiative on Sharing Avian Influenza Data to analyze the evolutionary and genetic features. RESULTS: We found that genetically different strains emerged in every epidemic in Shanghai, and most of the circulating H7N9 strains had affinity to human-type receptors, with the characteristics of high-virulence and low-pathogenic influenza viruses. Furthermore, our findings suggest that the Shanghai chicken strains are closely related to the HPAI H7N9 virus A/Guangdong/17SF003/2016, indicating that this viral strain is of avian origin and generated from the LPAI H7N9 viruses in Shanghai. The gradual decrease in H7N9 human infection in Shanghai was probably due to the control measures taken by the Shanghai government and the enhanced public awareness leading to a reduced risk of H7N9 virus infection. However, LPAI H7N9 viruses from poultry and environmental samples were continually detected in Shanghai across the epidemics, increasing the risk of new emerging H7N9 outbreaks. CONCLUSION: It is important to consistently obtain sufficient surveillance data and implement prevention measures against H7N9 viruses in Shanghai municipality.

Effects of water-borne copper on the Y-organ and content of 20-hydroxyecdysone in Eriocheir sinensis.

The effects of water-borne copper (Cu2+) (0.01, 0.10, 1.00, and 5.00 mg/L) on Y-organ (YO) microstructure and ultrastructure and on the content of 20-hydroxyecdysone (20-HE) in Eriocheir sinensis were studied using single gradient factor experiments. Results demonstrated that there was no significant difference in YO microstructure and ultrastructure when E. sinensis was exposed to 0.01, 0.10, and 1.00 mg/L water-borne Cu2+. However, exposure to 5.00 mg/L Cu2+ caused serious damage to the YO microstructure, with an obvious decrease in hemocoel and the disappearance of the basement membrane between cell fibers. In addition, there was an intumescence of cells around the hemocoel, the cell nucleus became enlarged or even rounded, and some cells around the hemocoel crumpled with only the cell nucleus isolated or even huddled in the hemocoel. Similarly, effects of water-borne Cu2+ on the YO ultrastructure were observed: obvious enlargement and rounding of the cell nucleus, high heterochromatinization of the nucleus chromatin, disappearance of the YO cell membrane in some severely damaged cells, and varied damage in cellular organelles, such as the disappearance of cristae in mitochondria and disruption of the nucleus membrane. The hemocyte nucleus showed considerable heterochromatinization. In addition, when the Cu2+ concentration increased, the content of 20-hydroxyecdysone (20-HE) decreased in all Cu2+ treatment groups (P < 0.05). Results of this study indicate that high water-borne Cu2+ concentration impaired the basic structure of YO, which is regarded as the secretion site of 20-HE, and hence decreased the content of 20-HE in hemolymph. Therefore, molting frequency as well as the growth of E. sinensis is inhibited when exposed to high concentrations of water-borne Cu2+.

Effective inhibition of porcine transmissible gastroenteritis virus replication in ST cells by shRNAs targeting RNA-dependent RNA polymerase gene.

Transmissible gastroenteritis virus (TGEV) is identified as one of the most important pathogenic agents during swine enteric infection, leading to high mortality in neonatal pigs and severe annual economic loss in swine-producing areas. Up to date, various vaccines developed against TGEV still need to be improved. To exploit the possibility of using RNA interference (RNAi) as a strategy against TGEV infection, two shRNA-expressing plasmids (pEGFP-U6/P1 and pEGFP-U6/P2) targeting the RNA-dependent RNA polymerase (RdRp) gene of TGEV were constructed and transfected into swine testicular (ST) cells. The cytopathic effect (CPE) and MTS assays demonstrated that both shRNAs were capable of protecting cells against TGEV invasion with very high specificity and efficiency. A real-time quantitative RT-PCR further confirmed that the amounts of viral RNAs in cell cultures pre-transfected with the two plasmids were reduced by 95.2% and up to 100%, respectively. Our results suggest that RNAi might be a promising new strategy against TGEV infection.