Abstract

Recent developments in molecular biology and metabolic engineering have resulted in a large increase in the number of strains that need to be tested, positioning high-throughput screening of microorganisms as an important step in bioprocess development. Scalability is crucial for performing reliable screening of microorganisms. Most of the scalability studies from microplate screening systems to controlled stirred-tank bioreactors have been performed so far with unicellular microorganisms. We have compared cultivation of industrially relevant oleaginous filamentous fungi and microalga in a Duetz-microtiter plate system to benchtop and pre-pilot bioreactors. Maximal glucose consumption rate, biomass concentration, lipid content of the biomass, biomass, and lipid yield values showed good scalability for Mucor circinelloides (less than 20% differences) and Mortierella alpina (less than 30% differences) filamentous fungi. Maximal glucose consumption and biomass production rates were identical for Crypthecodinium cohnii in microtiter plate and benchtop bioreactor. Most likely due to shear stress sensitivity of this microalga in stirred bioreactor, biomass concentration and lipid content of biomass were significantly higher in the microtiter plate system than in the benchtop bioreactor. Still, fermentation results obtained in the Duetz-microtiter plate system for Crypthecodinium cohnii are encouraging compared to what has been reported in literature. Good reproducibility (coefficient of variation less than 15% for biomass growth, glucose consumption, lipid content, and pH) were achieved in the Duetz-microtiter plate system for Mucor circinelloides and Crypthecodinium cohnii. Mortierella alpina cultivation reproducibility might be improved with inoculation optimization. In conclusion, we have presented suitability of the Duetz-microtiter plate system for the reproducible, scalable, and cost-efficient high-throughput screening of oleaginous microorganisms. Electronic supplementary material The online version of this article (10.1007/s00253-018-8920-x) contains supplementary material, which is available to authorized users. Document type: Article

Abstract

Objective: We studied which current fatal at-fault crashes would occur despite the most advanced current active safety devices (up to SAE level 2 of driving automation) and how frequent these crashes would be. Methods: We carried out a cross-sectional study of passenger cars that were first registered during the period 1st January 2010 to 31st December 2017 in Finland. To gain the true exposure for these cars, we accessed the national Vehicular and Driver Data Register to obtain the mileage information and the registration count for the study period of 2010-17. Similarly, we accessed the registry of Finnish road accident investigation teams and included all fatal at-fault crashes among the cars in our study for the same period. We used a real world reference technology for each active safety system in our analysis and chose one car brand as an example. This gave us exact system specifications and enabled testing the operation of the systems on the road. We performed field tests to gain further information on the precise operation of the safety systems in different operating conditions. Finally, we gathered all information on the studied active safety systems and analyzed the investigated at-fault fatal crashes case-by-case using our four level method. Results: Cars in our study were the primary party in 113 investigated fatal accidents during the years 2010-17. In 87 of the accidents, the leading cause of death was the injuries due to the crash, and these cases were classified as “unavoidable” (n = 58, 67 %), “avoidable” (n = 26, 30 %) or unsolved (n = 3, 3 %). Of the 58 “unavoidable” crashes 21 (36 %) were suicides, 21 (36%) involved active driver input which would have prevented the safety system operation, 15 (17 %) featured circumstances beyond the safety system performance and in one loss-of-control crash the driver had disabled the relevant safety system (electronic stability control). The registration years of the cars in our study (2010-17) totaled 3,772,864 and during this period, the cars travelled 75.9billion kilometers. The crash incidence of the “unavoidable” at-fault fatal crashes was 0.76-0.80 fatal crashes per billion kilometers and 15-16 fatal crashes per million registration years. Conclusions: We calculated a crash incidence for the “unavoidable” crashes which was 20–27% smaller than the observed crash rate of ESC-fitted passenger cars in our previous study. We concluded that suicides, active driver input until the crash, and challenging weather and road conditions are the most difficult factors for current active safety systems. Our analysis did not account for issues such as system usability or driver acceptance and therefore our results should be regarded as something that is currently theoretically achievable. However, the observed incidence is a good reference for automated driving development and the crash rate of automated cars. Peer reviewed