We report a 5.8-W deep-ultraviolet (DUV) laser obtained from frequency-quadrupling of an all-fiberized ytterbium-doped fiber (YDF) master oscillator power amplifier (MOPA). The MOPA system delivers 585 ps pulses at 1040 nm with a maximum available output power of 23.5 W for nonlinear frequency conversion. A lithium triborate (LBO) crystal and a beta barium borate (BBO) crystal are employed for second- and fourth-harmonic generation (FHG), respectively. At a repetition rate of 1.6 MHz, a maximum DUV output power of 5.8 W is obtained at 260 nm with a corresponding pulse energy of 3.6 μJ and maximum peak power of at least 6.9 kW. A 1μm-to-260nm conversion efficiency of 26.4% is achieved at a DUV output power of 5.8 W. To the best of our knowledge these results represent the highest-average-power fiberized-laser-pumped DUV laser, as well as the most efficient DUV generation based on BBO crystals to date. We further demonstrate application of the pulsed DUV laser in bacterial disinfection achieving an inactivation efficiency of 99.999% for E-coli bacteria at a DUV exposure of 7 mJ/cm².

Serial crystallography requires large numbers of microcrystals and robust strategies to rapidly apply substrates to initiate reactions in time-resolved studies. Here, we report the use of droplet miniaturization for the controlled production of uniform crystals, providing an avenue for controlled substrate addition and synchronous reaction initiation. The approach was evaluated using two enzymatic systems, yielding 3 µm crystals of lysozyme and 2 µm crystals of Pdx1, anArabidopsisenzyme involved in vitamin B6 biosynthesis. A seeding strategy was used to overcome the improbability of Pdx1 nucleation occurring with diminishing droplet volumes. Convection within droplets was exploited for rapid crystal mixing with ligands. Mixing times of <2 ms were achieved. Droplet microfluidics for crystal size engineering and rapid micromixing can be utilized to advance time-resolved serial crystallography.

Severe acute respiratory syndrome coronavirus 2 (SARS-Cov-2) has had a tremendous impact on humanity. Prevention of transmission by disinfection of surfaces and aerosols through a chemical-free method is highly desirable. Ultraviolet C (UVC) light is uniquely positioned to achieve inactivation of pathogens. We report the inactivation of SARS-CoV-2 virus by UVC radiation and explore its mechanisms. A dose of 50 mJ/cm² using a UVC laser at 266 nm achieved an inactivation efficiency of 99.89%, while infectious virions were undetectable at 75 mJ/cm² indicating >99.99% inactivation. Infection by SARS-CoV-2 involves viral entry mediated by the spike glycoprotein (S), and viral reproduction, reliant on translation of its genome. We demonstrate that UVC radiation damages ribonucleic acid (RNA) and provide in-depth characterization of UVC-induced damage of the S protein. We find that UVC severely impacts SARS-CoV- 2 spike protein's ability to bind human angiotensin-converting enzyme 2 (hACE2) and this correlates with loss of native protein conformation and aromatic amino acid integrity. This report has important implications for the design and development of rapid and effective disinfection systems against the SARS-CoV-2 virus and other pathogens.

Current methods for diagnosing acute and complex infections mostly rely on culture-based methods and, for biofilms, fluorescence in-situ hybridization. These techniques are labor-intensive and can take 2-4 days to return a test result, especially considering an extra culturing step required for the antibiotic susceptibility testing (AST). This places a significant burden on healthcare providers, delaying treatment and leading to adverse patient outcomes. Here, we report the complementary use of our newly developed multi-excitation Raman spectroscopy (ME-RS) method with whole-genome sequencing (WGS). Four WHO priority pathogens are AST phenotyped and their antimicrobial resistance (AMR) profile determined by WGS. On application of ME-RS method we find high correlation with the WGS characterization. Highly accurate classification based on the species (98.93%), wild-type/non-wild type (99.45%), and presence or absence of thick peptidoglycan layers in cell walls (100%), as well as at the individual strain level (99.29%). These results clearly demonstrate the potential of ME-RS as a rapid and first-stage tool for species, resistance and strain-level classification which can be followed up by WGS for confirmation. Such a workflow can facilitate efficient antimicrobial stewardship to handle and prevent the spread of AMR.

Oilfield systems are a multifaceted ecological niche, which consistently experiences microbiologically influenced corrosion. However, simulating the environmental conditions of an offshore system within the laboratory is notoriously difficult. A novel dual anaerobic biofilm reactor protocol allowed a complex mixed-species marine biofilm to be studied. Interestingly, electroactive corrosive bacteria and fermentative electroactive bacteria growth was supported within the biofilm microenvironment. Critically, the biotic condition exhibited pits with a greater average area, which is characteristic of microbiologically influenced corrosion. This research seeks to bridge the gap between experimental and real-world scenarios, ultimately enhancing the reliability of biofilm management strategies in the industry. IMPORTANCE: It is becoming more widely understood that any investigation of microbiologically influenced corrosion requires a multidisciplinary focus on multiple lines of evidence. Although there are numerous standards available to guide specific types of testing, there are none that focus on integrating biofilm testing. By developing a novel dual anaerobic reactor model to study biofilms, insights into the different abiotic and biotic corrosion mechanisms under relevant environmental conditions can be gained. Using multiple lines of evidence to gain a holistic understanding, more sustainable prevention and mitigation strategies can be designed. To our knowledge, this is the first time all these metrics have been combined in one unified approach. The overall aim of this paper was to explore recent advances in biofilm testing and corrosion research and provide recommendations for future standards being drafted. However, it is important to note that this article itself is not intending to serve as a standard.

Antimicrobial resistance (AMR) poses a global healthcare challenge, where overprescription of antibiotics contributes to its prevalence. We have developed a rapid multi-excitation Raman spectroscopy methodology (MX-Raman) that outperforms conventional Raman spectroscopy and enhances specificity. A support vector machine (SVM) model was used to identify 20 clinical isolates of Pseudomonas aeruginosa with an accuracy of 93% using MX-Raman. Antibiotic sensitivity profiles for tobramycin, ceftazidime, ciprofloxacin, and imipenem were generated for the bacterial strains and compared with their Raman spectral signatures using MX-Raman. The 20 clinical strains were distinguished according to AMR profiles. Nine models were assessed for AMR classification performance, and SVM performed best, classifying AMR profiles of each strain with 91–96% accuracy. These data provide the basis for a new rapid clinical diagnostic platform that could screen for bacterial infection and recommend effective antibiotic treatment ahead of confirmation by conventional techniques, improving clinical outcomes and reducing the spread of AMR.