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2018

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Sammendrag

Four lab scale biogas reactors fed with a substrate composition of ensiled fish waste and manure fixed at 13 and 87 vol %, respectively, were operated with HRTs of 20, 25, 30 and 40 days. Biogas process performance and stability were evaluated with regard to CH4 yields, NH4+ accumulation and abundance of NH4+-tolerant microorganisms. Process performance in the reactors operated at different HRTs were compared to process performance in reactors operated with constant HRT, fed with increased ratios of fish waste. The process performance and microbial dynamics were stable in reactors operated at constant amount of fish waste in the feed and with different HRTs. In the reactors added elevated ratios of fish waste, the concentration of NH4+ and abundance of NH4+-tolerant acetate oxidizing bacteria increased. The biogas process failed in these reactors simultaneously with an observed shift in microbial composition. In particular, the bacterium Tepidanaerobacter Acetatoxydans seemed to affect the biogas process stability. The hydrogenotrophic Methanomicrobiales increased in abundance in response to higher fish waste loading and NH4+ concentrations. This study showed that at a loading of 13% fish waste, it is possible to decrease the HRT from 30 to 20 days without markedly inhibiting the process stability.

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Dengue fever is a mosquito (Aedes aegypti) ‐transmitted viral disease that is endemic in more than 125 countries around the world. There are four serotypes of the dengue virus (DENV 1‐4) and a safe and effective dengue vaccine must provide protection against all four serotypes. To date, the first vaccine, Dengvaxia (CYD‐TDV), is available after many decades’ efforts, but only has moderate efficacy. More effective and affordable vaccines are hence required. Plants offer promising vaccine production platforms and food crops offer additional advantages for the production of edible human and animal vaccines, thus eliminating the need for expensive fermentation, purification, cold storage and sterile delivery. Oral vaccines can elicit humoral and cellular immunity via both the mucosal and humoral immune systems. Here, we report the production of tetravalent EDIII antigen (EDIII‐1‐4) in stably transformed lettuce chloroplasts. Transplastomic EDIII‐1‐4‐expressing lettuce lines were obtained and homoplasmy was verified by Southern blot analysis. Expression of EDIII‐1‐4 antigens was demonstrated by immunoblotting, with the EDIII‐1‐4 antigen accumulating to 3.45% of the total protein content. Immunological assays in rabbits showed immunogenicity of EDIII‐1‐4. Our in vitro gastrointestinal digestion analysis revealed that EDIII‐1‐4 antigens are well protected when passing through the oral and gastric digestion phases but underwent degradation during the intestinal phase. Our results demonstrate that lettuce chloroplast engineering is a promising approach for future production of an affordable oral dengue vaccine.