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Chicken coccidiosis caused by Eimeria spp. can occur on almost all poultry farms, causing huge economic losses to the industry. Genetically manipulated Eimeria parasites as a vaccine vector to deliver viral antigens have been reported. In our preliminary study, transgenic E. acervulina expressing a VP2 gene (Ea-VP2) of the infectious bursal disease virus (IBDV) demonstrated partial protection against IBDV infection. To enhance immune responses, we aimed to increase the VP2 gene copy number in transgenic E. acervulina. In this study, we used a novel plasmid vector carrying a VP2 gene fused with three flag tags and a red fluorescent reporter gene (mCherry). The vector was introduced into Ea-VP2 sporozoites through nucleofection, leading to the generation of Ea-2VP2. Subsequent analysis revealed a notable escalation in the fluorescent rate, increasing from 0.11 to 95.1% following four consecutive passages facilitated by fluorescent-activated cell sorting. Verification via PCR, Western blot, and immunofluorescence confirmed the successful construction of the Ea-2VP2 population. Despite lower fecundity compared to wild-type E. acervulina, Ea-2VP2 maintained immunogenicity. Our research effectively created a transgenic E. acervulina strain transfected sequentially with two copies of the VP2 gene from IBDV. This modification resulted in an increased humoral immune response after primary immunization in chickens. Additionally, it demonstrated a degree of protection within the bursa against IBDV infection. Future studies will focus on further enhancing immune response levels. [ABSTRACT FROM AUTHOR]

Streeck might argue that there should be no global economy, but many states would continue to participate in one, there will still be global pollution and other issues that simply cannot be confronted by individual states, especially small ones. Outright opponents of globalization, like Streeck, need to recognize that, if the process were to be reversed and a serious deglobalization to take place, income levels across the newly industrializing lands would fall, leading not only to increased poverty there, but also a loss of trade for existing wealthy countries. One of the many other reasons for Streeck's hostility to the European Union and to European integration in general is his claim that small and medium-sized states are frequently better governed and economically more effective than large ones. These other countries, Y and Z, retaliate by imposing tariffs on a successful export of country X. The negative effects of tariffs would be experienced fairly quickly by country X, while it would take some time, perhaps years, before its motor industry had revived. [Extracted from the article]