The toxicity of waste printed circuit boards (PCBs) to bacteria was considered as the major limitation in bioleaching of copper from PCBs. To reduce the toxicity of PCBs, copper extraction from PCBs was investigated using bacteria-free cultural supernatant from some metallurgical microbial consortium, whose predominant organisms were Leptospirillum ferriphilum and Sulfobacillus thermosulfidooxidans. About 100% copper was recovered in 2 h from 5 g/L PCBs by bacteria-free cultural supernatant. The result indicated that the indirect non-contact mechanism was the predominant mechanism in bioleaching of copper from PCBs. It was not necessary for bacteria to exist in copper extraction. In addition, the role of bacteria was most likely to regenerate Fe3+ as an oxidant. Furthermore, the biooxidation of Fe2+ to Fe3+ was determined as the rate-limited step in bioleaching of copper from PCBs. In addition, the biooxidation activity of bacteria would be strongly inhibited by the toxicity of PCBs. The separation of bacteria from the PCBs probably was the optimum way to avoid the negative effect of PCBs. Accordingly, a biooxidation–leaching–separation cycle was designed to avoid the toxicity of PCBs. Eventually, 93.4% of copper was recovered in total from 100 g/L PCB concentrates in 9 days.

Influenza vaccines for H7N9 subtype have shown low immunogenicity in human clinical trials. Using novel adjuvants might represent the optimal available option in vaccine development. In this study, we demonstrated that the using of the STING agonist cGAMP as a mucosal adjuvant is effective in enhancing humoral, cellular and mucosal immune responses of whole virus, inactivated H7N9 vaccine in mice. A single dose of immunization was able to completely protect mice against a high lethal doses of homologous virus challenge with an significant dose-sparing effect. We also found that intranasal co-administration of H7N9 vaccine with cGAMP could provide effective cross protection against H1N1, H3N2, and H9N2 influenza virus. Furthermore, cGAMP induced significantly higher nucleoprotein specific CD4⁺ and CD8⁺ T cells responses in immunized mice, as well as upregulated the IFN-γ and Granzyme B expression in the lung tissue of mice in the early stages post a heterosubtypic virus challenge. These results indicated that STING agonist cGAMP was expected to be an effective mucosal immune adjuvant for pre-pandemic vaccines such as H7N9 vaccines, and the cGAMP combined nasal inactivated influenza vaccine will also be a promising strategy for development of broad-spectrum influenza vaccines.

Development of serum-free suspension cell culture processes is very important for influenza vaccine production. Previously, we developed a MDCK suspension cell line in a serum-free medium. In the present study, the growth kinetics of suspension MDCK cells and influenza virus production in the serum-free medium were investigated, in comparison with those of adherent MDCK cells in both serum-containing and serum-free medium. It was found that the serum-free medium supported the stable subculture and growth of both adherent and suspension cells. In batch culture, for both cell lines, the growth kinetics in the serum-free medium was comparable with those in the serum-containing medium and a commercialized serum-free medium. In the serum-free medium, peak viable cell density (VCD), haemagglutinin (HA) and median tissue culture infective dose (TCID₅₀) titers of the two cell lines reached 4.51×10⁶ cells/mL, 2.94Log₁₀(HAU/50 μL) and 8.49Log₁₀(virions/mL), and 5.97×10⁶ cells/mL, 3.88Log₁₀(HAU/50 μL), and 10.34Log₁₀(virions/mL), respectively. While virus yield of adherent cells in the serum-free medium was similar to that in the serum-containing medium, suspension culture in the serum-free medium showed a higher virus yield than adherent cells in the serum-containing medium and suspension cells in the commercialized serum-free medium. However, the percentage of infectious viruses was lower for suspension culture in the serum-free medium. These results demonstrate the great potential of this suspension MDCK cell line in serum-free medium for influenza vaccine production and further improvements are warranted.