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Extracellular vesicles (EVs) function as potent mediators of intercellular communication for many in vivo processes, contributing to both health and disease related conditions. Given their biological origins and diverse functionality from correspondingly unique "cargo" compositions, both endogenous and modified EVs are garnering attention as promising therapeutic modalities and vehicles for targeted therapeutic delivery applications. Their diversity in composition, however, has revealed a significant need for more comprehensive analytical-based characterization methods, and manufacturing processes that are consistent and scalable. In this review, we explore the dynamic landscape of EV research and development efforts, ranging from novel isolation approaches, to their analytical assessment through novel characterization techniques, and to their production by industrial-scale manufacturing process considerations. Expanding the horizon of these topics to EVs for in-human applications, we underscore the need for stringent development and adherence to Good Manufacturing Practice (GMP) guidelines. Wherein, the intricate interplay of raw materials, production in bioreactors, and isolation practices, along with analytical assessments compliant with the Minimal Information for Studies of Extracellular Vesicles (MISEV) guidelines, in conjunction with reference standard materials, collectively pave the way for standardized and consistent GMP production processes.
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The regrowth and subsequent exposure of opportunistic pathogens (OPs) whilst reopening buildings that have been locked down due to the stay-at-home restrictions to limit the spread of COVID-19, is a public health concern. To better understand such microbiological risks due to lowered occupancy and water demand in buildings, first and post-flush water samples (n = 48) were sampled from 24 drinking water outlets from eight university buildings in two campuses (urban and rural), with various end-user occupancies. Both campuses were served with chlorinated water originating from a single drinking water distribution system in South-East Queensland, situated 14 km apart, where the rural campus had lower chlorine residuals. Culture-dependent and culture-independent methods (such as flow cytometry, qPCR and 16S rRNA gene amplicon sequencing) were used concurrently to comprehensively characterise the OPs of interest (Legionella spp., Pseudomonas aeruginosa, and nontuberculous mycobacteria (NTM)) and the premise plumbing microbiome. Results showed that buildings with extended levels of stagnation had higher and diverse levels of microbial growth, as observed in taxonomic structure and composition of the microbial communities. NTM were ubiquitous in all the outlets sampled, regardless of campus or end-user occupancy of the buildings. qPCR and culture demonstrated prevalent and higher concentrations of NTM in buildings (averaging 3.25 log10[estimated genomic copies/mL]) with extended stagnation in the urban campus. Furthermore, flushing the outlets for 30 minutes restored residual and total chlorine, and subsequently decreased the levels of Legionella by a reduction of 1 log. However, this approach was insufficient to restore total and residual chlorine levels for the outlets in the rural campus, where both Legionella and NTM levels detected by qPCR remained unchanged, regardless of building occupancy. Our findings highlight that regular monitoring of operational parameters such as residual chlorine levels, and the implementation of water risk management plans are important for non-healthcare public buildings, as the levels of OPs in these environments are typically not assessed.
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Biotrickling filter (BTF) is a widely applied bioreactor for odour abatement in sewer networks. The trickling strategy is vital for maintaining a sound operation of BTF. This study employed a lab-scale BTF packed with granular activated carbon at a short empty bed residence time of 6 s and pH 1-2 to evaluate different trickling strategies, i.e., continuous trickling (different velocities) and intermittent trickling (different trickling intervals), in terms of the removal of hydrogen sulfide (H2S), bed pressure drop, H2S oxidation products and microbial community. The H2S removal performance decreased with the trickling velocity (â¼3.6 m/h) in BTF. In addition, three intermittent trickling strategies, i.e., 10-min trickling per 24 h, 8 h, and 2 h, were investigated. The H2S elimination capacity deteriorated after about 2 weeks under both 10-min trickling per 24 h and 8 h. For both intermittent (10-min trickling per 2 h) and continuous trickling, the BTF exhibited nearly 100 % H2S removal for inlet H2S concentrations<100 ppmv, but intermittent BTF showed better removal performance than continuous trickling when inlet H2S increased to 120-190 ppmv. Furthermore, the bed pressure drops were 333 and 3888 Pa/m for non-trickling and trickling periods, respectively, which makes intermittent BTF save 83 % energy consumption of the blower compared with continuous tirckling. However, intermittent BTF exhibited transient H2S breakthrough (<1 ppmv) during trickling periods. Moreover, elemental sulfur and sulfate were major products of H2S oxidation and Acidithiobacillus was the dominant genus in both intermittent and continuous trickling BTF. A mathematical model was calibrated for the intermittent BTF and a sensitivity analysis was performed on the model. It shows mass transfer parameters determine the H2S removal. Overall, intermittent trickling strategy is promising for improving odour abatement performance and reducing the operating cost of the BTF.
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Sulfuro de Hidrógeno , Microbiota , Biodegradación Ambiental , Reactores Biológicos , Filtración , AzufreRESUMEN
The premise plumbing portion of drinking water distribution systems (DWDS) has several characteristics that may favor microbial growth in the form of biofilms. These microbial communities are implicated as infectious sources for the spread of opportunistic waterborne pathogens by supporting their complex ecology and transmission through DWDS outlets to susceptible individuals. However, there is limited understanding of the drinking water biofilms in real premise plumbing networks due to challenges with accessibility. Using a combination of culture-dependent and culture-independent approaches, this study comprehensively characterized the premise plumbing microbiome of a 50-year-old university building, inclusive of water and biofilm samples. Microbial diversity in the water samples were more taxonomically diverse in comparison to the mature drinking water biofilms, which were dominated with biofilm-formers and opportunistic pathogens, such as Mycobacterium spp. A model opportunistic pathogen, Legionella spp., was only detectable in water samples using quantitative PCR but could not be detected in any of the drinking water biofilms using either qPCR or culture-dependent approaches, highlighting the limitations of detection methods in these environments. This study presents preliminary findings on the microbial dynamics and complexity in premise plumbing networks, which may support public health management and the development of strategies to eliminate microbial risks to human health.