Mathematical Modeling for Evaluating Inherent Parameters Affecting UVC Decontamination of Indicator Bacteria.

UV light is a tool associated with the denaturation of cellular components, DNA damage, and cell disruption. UV treatment is widely used in the decontamination process; however, predicting a sufficient UV dose by using traditional methods is doubtful. In this study, an in-house UVC apparatus was designed to investigate the process of the inactivation of five indicator bacteria when the initial cell concentrations and irradiation intensities varied. Both linear and nonlinear mathematical models were applied to predict the inactivation kinetics. In comparison with the Weibull and modified Chick-Watson models, the Chick-Watson model provided a good fit of the experimental data for five bacteria, Escherichia coli, Klebsiella pneumoniae, Staphylococcus aureus, Streptococcus faecalis, and Bacillus subtilis. The specific death rate (kd) significantly increased when the irradiation intensity (I) increased from 1.41 W/m2 to 3.02 W/m2 and 4.83 W/m2 (P < 0.05). Statistical analysis revealed no significant difference in the kd values among the groups of tested Gram-positive bacteria, Gram-negative bacteria, and B. subtilis spores, but the kd values differed among groups (P < 0.05). The death rate coefficient (k) varied from species to species. The k values of the tested Gram-positive bacteria were higher than those of the Gram-negative bacteria. The thick peptidoglycan layer in the Gram-positive membrane was responsible for UVC resistance. The high guanine-cytosine (GC) content in bacteria also contributed to UV resistance due to the less photoreactive sites on the nucleotides. This investigation provides a good understanding of bacterial inactivation induced by UVC treatment. IMPORTANCE Prevention and control measures for microbial pathogens have attracted worldwide attention due to the recent coronavirus disease 2019 pandemic. UV treatments are used as a commercial control to prevent microbial contamination in diverse applications. Microorganisms exhibit different UV sensitivities, which are often measured by the UV doses required for decreasing the number of microbial contaminants in the logarithmic order. The maximum efficacy of UV is usually observed at 254 nm (residing in the UVC range of the light spectrum). UV technology is a nonthermal physical decontamination measure that does not require any chemicals and consumes low levels of energy while leaving insignificant amounts of chemical residues or toxic compounds. Therefore, obtaining the microbial death kinetics and their intrinsic parameters provided in this study together with the UV photoreaction rate enables advancement in the design of UV treatment systems.

Bactericidal effect of ultraviolet C light-emitting diodes: Optimization of efficacy toward foodborne pathogens in water.

The elimination of bacterial pathogens from water using ultraviolet C light-emitting diodes (UVC-LEDs) is a critical technology in terms of hygiene and sanitation. This technology has several advantages, such as low energy consumption, no heating requirements, and high effectiveness. Although several studies have reported the bactericidal effect of UVC-LEDs, little information is available on their bactericidal effect on water reservoirs contaminated with microorganisms. Therefore, the aim of this study was to optimize the bactericidal effects of UVC-LED irradiation, particularly at a wavelength of 278 nm, against major foodborne gram-positive and gram-negative pathogenic bacteria, such as Escherichia coli, Staphylococcus aureus, Bacillus cereus, Salmonella Typhimurium, and Listeria monocytogenes. The efficiency of the bactericidal effect of UVC-LED irradiation was determined based on three variables: exposure time (A, 0-60 min), stirring speed (B, 0-100 rpm), and volume of water (C, 400-1200 mL). To optimize the conditions, the operation of the designed model and results analysis were carried out using Box-Behnken design (BBD) and response surface method (RSM). The final conditions optimized for an effective bactericidal activity included a 60 min exposure time, a 100 rpm stirring speed, and 400 mL of liquid volume. Furthermore, the validation of the optimized model using the predicted values was calculated by the program, which was conducted by matching the actual values within standard deviations. The present study revealed that the optimization of a UVC-LED irradiation model is a promising approach for effectively controlling the contamination of water reservoirs by bacterial pathogens.

The antimicrobial effect of 400 nm femtosecond laser and silver nanoparticles on gram-positive and gram-negative bacteria.

Silver nanoparticles are well-known for their antimicrobial effect. However, they are potentially toxic in high doses. We explored the possibility of enhancing the bactericidal effect of low concentrations of silver nanoparticles with blue light femtosecond laser irradiation, since such concentrations are less toxic. The growth dynamics of Pseudomonas aeruginosa, Listeria monocytogenes and methicillin-resistant Staphylococcus aureus grown in pre-synthesized silver nanoparticles were measured with or without pre-irradiation with 50 mW and 400 nm femtosecond laser irradiation. With each bacterium, combined treatment with laser and silver nanoparticles significantly reduced bacterial growth, indicating that this form of treatment could be beneficial in the ongoing efforts to reduce the deleterious effects of antibiotic resistant Gram-positive and Gram-negative bacteria. The combined treatment was more antimicrobial than treatment with silver nanoparticles alone or photo-irradiation alone. P. aeruginosa and L. monocytogenes were more susceptible to the bactericidal effects of silver nanoparticles, and the combination of laser treatment and silver nanoparticles than MRSA.

Synergistic bactericidal effects of pairs of photosensitizer molecules activated by ultraviolet A light against bacteria in plasma.

BACKGROUND: The current approach to reducing bacterial contamination in blood transfusion products is through detection or pathogen reduction methods, some of which utilize ultraviolet (UV) light photosensitizers. A small number of photosensitizers are being used as single agents in combination with UV light, but their efficacy can be limited against some pathogens. Benzophenone (BP) and vitamins B1, B6, and K3 have been identified as effective UVA photosensitizers for inactivation of bacteria. We evaluated whether combining pairs of photosensitizers in this group would have synergistic bactericidal effects on Gram-negative and Gram-positive bacteria. STUDY DESIGN AND METHODS: Bacteria species of Escherichia coli, Bacillus cereus, Staphylococcus aureus, and Klebsiella pneumoniae were mixed with 0 to 100 mM concentrations of photosensitizers and exposed to UVA irradiation at 18 J/cm2 to assess their bactericidal effects. RESULTS: Single photosensitizers irradiated with UVA produced a range of bactericidal activity. When combined in pairs, all demonstrated some synergistic bactericidal effects with up to 4-log reduction above the sum of activities of individual molecules in the pair against bacteria in plasma. Photosensitizer pairs with BP had the highest synergism across all bacteria. With vitamin K3 in the pair, synergism was evident for Gram-positive but not for Gram-negative bacteria. Vitamin B1 and vitamin B6 had the least synergism. These results indicate that a combination approach with multiple photosensitizers may extend effectiveness of pathogen reduction in plasma. CONCLUSIONS: Combining photosensitizers in pathogen reduction methods could improve bactericidal efficacy and lead to use of lower concentrations of photosensitizers to reduce toxicities and unwanted side effects.

Efficient Photodynamic Killing of Gram-Positive Bacteria by Synthetic Curcuminoids.

In our previous study, we have demonstrated that curcumin can efficiently kill the anaerobic bacterium Propionibacterium acnes by irradiation with low-dose blue light. The curcuminoids present in natural plant turmeric mainly include curcumin, demethoxycurcumin, and bisdemethoxycurcumin. However, only curcumin is commercially available. Eighteen different curcumin analogs, including demethoxycurcumin and bisdemethoxycurcumin, were synthesized in this study. Their antibacterial activity against Gram-positive aerobic bacteria Staphylococcus aureus and Staphylococcus epidermidis was investigated using the photodynamic inactivation method. Among the three compounds in turmeric, curcumin activity is the weakest, and bisdemethoxycurcumin possesses the strongest activity. However, two synthetic compounds, (1E,6E)-1,7-bis(5-methylthiophen-2-yl)hepta-1,6-diene-3,5-dione and (1E,6E)-1,7-di(thiophen-2-yl)hepta-1,6-diene-3,5-dione, possess the best antibacterial activity among all compounds examined in this study. Their chemical stability is also better than that of bisdemethoxycurcumin, and thus has potential for future clinical applications.

Photodynamic Inactivation of Bacteria with Porphyrin Derivatives: Effect of Charge, Lipophilicity, ROS Generation, and Cellular Uptake on Their Biological Activity In Vitro.

Resistance of microorganisms to antibiotics has led to research on various therapeutic strategies with different mechanisms of action, including photodynamic inactivation (PDI). In this work, we evaluated a cationic, neutral, and anionic meso-tetraphenylporphyrin derivative's ability to inactivate the Gram-negative and Gram-positive bacteria in a planktonic suspension under blue light irradiation. The spectroscopic, physicochemical, redox properties, as well as reactive oxygen species (ROS) generation capacity by a set of photosensitizers varying in lipophilicity were investigated. The theoretical calculations were performed to explain the distribution of the molecular charges in the evaluated compounds. Moreover, logP partition coefficients, cellular uptake, and phototoxicity of the photosensitizers towards bacteria were determined. The role of a specific microbial efflux pump inhibitor, verapamil hydrochloride, in PDI was also studied. The results showed that E. coli exhibited higher resistance to PDI than S. aureus (3-5 logs) with low light doses (1-10 J/cm2). In turn, the prolongation of irradiation (up to 100 J/cm2) remarkably improved the inactivation of pathogens (up to 7 logs) and revealed the importance of photosensitizer photostability. The PDI potentiation occurs after the addition of KI (more than 3 logs extra killing). Verapamil increased the uptake of photosensitizers (especially in E. coli) due to efflux pump inhibition. This effect suggests that PDI is mediated by ROS, the electrostatic charge interaction, and the efflux of photosensitizers (PSs) regulated by multidrug-resistance (MDR) systems. Thus, MDR inhibition combined with PDI gives opportunities to treat more resistant bacteria.

Effects of a novel gel containing 5-aminolevulinic acid and red LED against bacteria involved in peri-implantitis and other oral infections.

Antibiotic resistance is a major public health problem worldwide and the finding of alternative methods for eliminating bacteria is one of the prerogatives of medical research. The indiscriminate use of antibiotics in dentistry, especially for the treatment of peri-implantitis, could lead to superinfections. Alternative methods, like photodynamic therapy mediated by the use of aminolevulinic acid and a red light has been largely described, especially in dentistry, but results were encouraging against Gram-positive bacteria, but limited against Gram-negative. The effectiveness of photodynamic therapy mediated by a novel product containing aminolevulinic acid, Aladent (ALAD) has been tested in this in vitro study, against different types of bacteria particularly involved in the infections of the oral cavity and peri-implantitis. The novelty of ALAD is the marked hydrophilicity that should increase the passage of the molecule through the membrane pores of Gram-negative bacteria. Considering the novelty of the product a preliminary experiment permitted to test the effectiveness against Enterococcus faecalis after 1 h of ALAD incubation at different concentrations, with or without different timings of LED irradiation. The count of CFUs and the live/dead observation with fluorescent microscopy showed a significant reduction and killing of bacterium. Then, in the second stage, that could meet the necessity of effectiveness and the clinician's requests to reduce the timing of treatment, ALAD, with and without irradiation, was tested on Staphylococcus aureus, Enterococcus faecalis, Escherichia coli, Veillonella parvula and Porphyromonas gingivalis. In particular, the efficiency of different concentrations of the product after a 25 min incubation was tested with and without the adjunctive LED irradiation for 5 min. A slight ALAD bactericidal effect was reported for all bacteria, also without LED irradiation, however, the most effective treatment was 25 min of 50% ALAD incubation followed by 5 min of a red LED. The in vitro tests demonstrated that ALAD gel with LED irradiation exerts a potent antibacterial activity on different bacteria, both Gram-positive and Gram-negative.

Thermal-Disrupting Interface Mitigates Intercellular Cohesion Loss for Accurate Topical Antibacterial Therapy.

Bacterial infections remain a leading threat to global health because of the misuse of antibiotics and the rise in drug-resistant pathogens. Although several strategies such as photothermal therapy and magneto-thermal therapy can suppress bacterial infections, excessive heat often damages host cells and lengthens the healing time. Here, a localized thermal managing strategy, thermal-disrupting interface induced mitigation (TRIM), is reported, to minimize intercellular cohesion loss for accurate antibacterial therapy. The TRIM dressing film is composed of alternative microscale arrangement of heat-responsive hydrogel regions and mechanical support regions, which enables the surface microtopography to have a significant effect on disrupting bacterial colonization upon infrared irradiation. The regulation of the interfacial contact to the attached skin confines the produced heat and minimizes the risk of skin damage during thermoablation. Quantitative mechanobiology studies demonstrate the TRIM dressing film with a critical dimension for surface features plays a critical role in maintaining intercellular cohesion of the epidermis during photothermal therapy. Finally, endowing wound dressing with the TRIM effect via in vivo studies in S. aureus infected mice demonstrates a promising strategy for mitigating the side effects of photothermal therapy against a wide spectrum of bacterial infections, promoting future biointerface design for antibacterial therapy.

Tribenzoporphyrazines with dendrimeric peripheral substituents and their promising photocytotoxic activity against Staphylococcus aureus.

Infectious diseases constitute a serious problem for human health and life. Although many bacterial and fungal infections can be successfully cured by commonly used antibiotics, a new threat emerges in the form of microbial resistance. For this reason, researchers try to find not only new active pharmaceutical ingredients for conventional antibiotherapy but also try to develop new strategies of microbial inactivation. Photodynamic antimicrobial chemotherapy, which relies on reactive oxygen species generated in situ in the presence of a photosensitizer and with the light of an appropriate wavelength, is one of them. Porphyrazines have been considered as potential photosensitizers for anticancer and antimicrobial photodynamic therapy. In this study, three tribenzoporphyrazines with dendrimeric peripheral substituents were subjected to in vitro antimicrobial photocytotoxicity study. One magnesium(II) tribenzoporphyrazine with peripheral 3,5-bis(3,5-dimethoxybenzyloxy)benzylsulfanyl substituents was synthesized and subjected to physicochemical characterization using NMR, UV-Vis, and mass spectrometry techniques. In photochemical studies this molecule revealed moderate singlet oxygen generation ability (ΦΔDMF = 0.12, ΦΔDMSO = 0.13). The other two magnesium(II) tribenzoporphyrazines applied in the biological study were 4-[3,5-di(hydroxymethyl)phenoxy]butylsulfanyl-substituted tribenzoporphyrazine and 4-[3,5-bis(benzyloxy)benzyloxy]phenyl-substituted tribenzopyrazinoporphyrazine. For the assessment, three microbial strains were chosen: Gram-positive bacteria Staphylococcus aureus ATCC 25923, Gram-negative bacteria Escherichia coli ATCC 25922, and fungal strain Candida albicans ATCC 10231. Very high activity against Staphylococcus aureus at low 10-6 M concentration was recorded for magnesium(II) tribenzoporphyrazines with peripheral 3,5-bis(3,5-dimethoxybenzyloxy)benzylsulfanyl and 4-[3,5-di(hydroxymethyl)phenoxy]butylsulfanyl substituents with calculated log reductions of 4.4 and 4.8, respectively. It is worth noting that magnesium(II) tribenzoporphyrazine with 4-[3,5-di(hydroxymethyl)phenoxy]butylsulfanyl substituents revealed also 3.2 log reduction in bacterial growth at the concentration 10-7 M.

Hypocrellin B-Mediated Photodynamic Inactivation of Gram-Positive Antibiotic-Resistant Bacteria: An In Vitro Study.

Background: The search for alternative therapeutics against antibiotic-resistant bacteria is highly desirable. A promising approach is photodynamic antimicrobial chemotherapy. Objective: This work evaluated the photodynamic inactivation (PDI) efficacy of hypocrellin B (HB) on Gram-positive antibiotic-resistant bacteria. Methods: PDI efficacy of HB on Gram-positive standard and antibiotic-resistant Staphylococcus aureus, Enterococcus faecalis, and Streptococcus pneumonia and Gram-negative Escherichia coli and Klebsiella pneumoniae was assessed. HB photoactivity on biofilms formed by the Gram-positive bacteria and its cytotoxicity on mammalian CT26 cells were also investigated. Results: HB showed no obvious dark toxicity, but provided concentration-dependent inactivation of bacteria and mammalian cells. After irradiation with 72 J/cm2 light, 100 µM of HB achieved about 7 log10 reductions in bacterial survival of Gram-positive strains, but yielded only 2 log10 reductions in bacterial survival of Gram-negative strains. Gram-positive bacteria were as susceptible to PDI in biofilms as in planktonic suspensions, but the efficacy was attenuated. Conclusions: The results suggested that HB could serve as a potential antibacterial photosensitizer against Gram-positive antibiotic-resistant bacteria.

Evaluation of Structure-Function Relationships of Aggregation-Induced Emission Luminogens for Simultaneous Dual Applications of Specific Discrimination and Efficient Photodynamic Killing of Gram-Positive Bacteria.

Bacterial infectious diseases, especially those caused by Gram-positive bacteria, have been seriously threatening human health. Preparation of a multifunctional system bearing both rapid bacterial differentiation and effective antibacterial effects is highly in demand, but remains a severe challenge. Herein, we rationally designed and successfully developed a sequence of aggregation-induced emission luminogens (AIEgens) with orderly enhanced D-A strength. Evaluation of structure-function relationships reveals that AIEgens having intrinsic positive charge and proper ClogP value are able to stain Gram-positive bacteria. Meanwhile, one of the presented AIEgens (TTPy) can generate reactive oxygen species (ROS) in extraordinarily high efficiency under white light irradiation due to the smaller singlet-triplet energy gap. Thanks to the NIR emission, excellent specificity to Gram-positive bacteria, and effective ROS generation efficiency, TTPy has been proved to perform well in selective photodynamic killing of Gram-positive bacteria in vitro, such as S. aureus and S. epidermidis, even in S. aureus-infected rat wounds.

Bacteria-induced aggregation of bioorthogonal gold nanoparticles for SERS imaging and enhanced photothermal ablation of Gram-positive bacteria.

The challenge in antimicrobial photothermal therapy (PTT) is to develop strategies for decreasing the damage to cells and increasing the antibacterial efficiency. Herein, we report a novel theranostic strategy based on bacteria-induced gold nanoparticle (GNP) aggregation, in which GNPs in situ aggregated on the bacterial surface via specific targeting of vancomycin and bioorthogonal cycloaddition. Plasmonic coupling between adjacent GNPs exhibited a strong "hot spot" effect, enabling effective surface enhanced Raman scattering (SERS) imaging of bacterial pathogens. More importantly, in situ aggregation of GNPs showed strong NIR adsorption and high photothermal conversion, allowing enhanced photokilling activity against Gram-positive bacteria. In the absence of bacterial strains, GNPs were dispersed and showed a very low photothermal effect, minimizing the side effects towards surrounding healthy tissues. Given the above advantages, the bioorthogonal theranostic strategy developed in this study may find potential applications in treating bacterial infection and even multidrug-resistant bacteria.

Comparison of Anti-Microbial Effects of Low-Level Laser Irradiation and Microwave Diathermy on Gram-Positive and Gram-Negative Bacteria in an In Vitro Model.

Background and Objectives: The aim of this study was to compare the effects of low-level laser therapy and continuous microwave diathermy on the growth of Gram-negative and Gram-positive bacteria and to establish their efficacy as an alternative therapeutic modality. MATERIALS AND METHODS: Laser fluence of 13 Joules (J)/cm2, 18 J/cm2 and 30 J/cm2 were used against several bacterial strains. Microwave dosages of 25, 50 and 100 watts (W) were used, respectively. RESULTS: A significant difference between the three groups was observed using repeated analysis of variance (RANOVA) (F value: 0.74, and p value: 0.001). The Greenhouse-Geisser correction (GG) revealed significant results for laser irradiation alone. However, effect size calculation showed effects with microwave diathermy as well as laser fluence. CONCLUSIONS: Low-level laser therapy appears to be an effective modality of treatment when compared with continuous microwave diathermy on the Gram-negative and the Gram-positive bacterial strains tested. Microwave diathermy revealed large and medium effects on the bacterial cell counts with dominant effects on Gram-negative strains.

Characterizing the Antimicrobial Properties of 405 nm Light and the Corning® Light-Diffusing Fiber Delivery System.

BACKGROUND AND OBJECTIVES: Hospital-acquired infections (HAIs) and multidrug resistant bacteria pose a significant threat to the U.S. healthcare system. With a dearth of new antibiotic approvals, novel antimicrobial strategies are required to help solve this problem. Violet-blue visible light (400-470 nm) has been shown to elicit strong antimicrobial effects toward many pathogens, including representatives of the ESKAPE bacterial pathogens, which have a high propensity to cause HAIs. However, phototherapeutic solutions to prevention or treating infections are currently limited by efficient and nonobtrusive light-delivery mechanisms. STUDY DESIGN/MATERIALS AND METHODS: Here, we investigate the in vitro antimicrobial properties of flexible Corning® light-diffusing fiber (LDF) toward members of the ESKAPE pathogens in a variety of growth states and in the context of biological materials. Bacteria were grown on agar surfaces, in liquid culture and on abiotic surfaces. We also explored the effects of 405 nm light within the presence of lung surfactant, human serum, and on eukaryotic cells. Pathogens tested include Enterococcus spp, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa, Enterobacter spp., Staphylococcus epidermidis, Streptococcus pyogenes, Candida albicans, and Escherichia coli. RESULTS: Overall, the LDF delivery of 405 nm violet-blue light exerted a significant degree of microbicidal activity against a wide range of pathogens under diverse experimental conditions. CONCLUSIONS: The results exemplify the fiber's promise as a non-traditional approach for the prevention and/or therapeutic intervention of HAIs. Lasers Surg. Med. © 2019 The Authors. Lasers in Surgery and Medicine Published by Wiley Periodicals, Inc.

Photocatalytic decomposition effect of erbium doped cerium oxide nanostructures driven by visible light irradiation: Investigation of cytotoxicity, antibacterial growth inhibition using catalyst.

Cerium (IV) oxide (CeO2) is the most accessible noble rare earth metal oxide for the excitation of the excitons by light-harvesting performance. The present work is focused on Erbium doped ceria nanoparticles that were beneficially obtained by hydrothermal method from cerium nitrate and Erbium nitrate as precursors for decomposition of Rhodamine-B (RhB) dye in the polluted waste water removed from the industries. Dye removal efficiency of the catalyst was found to be nearly ~94%. The structural phases, functional groups and the transitions are identified with the help of various techniques. XRD pattern determines the development of cubic phase with the particle size is 20 nm. Highly crystalline nature of as-synthesized nanomaterials with an average diameter of 35 nm was investigated by HRSEM. The crystalline size, shape and textural morphology, of the Erbium doped ceria nanostructures were analysed by HRTEM. Our results suggest, that the concentration of OH- ion determines the lattice constants and oxygen vacancy in the nanostructures which stimulate the probability of photocatalytic decomposition effect of organic pollutants, due to synergistic approach. In this context, both unhydrolyzed things and their swiftly drip from deceased or scratched cells with conceded membranes, even when the cells embrace some are outstanding attention. Although, the loss of viable cells also depends on epithelial cell dynamically conceal of numerous molar matrix.

Inactivation dynamics of 222 nm krypton-chlorine excilamp irradiation on Gram-positive and Gram-negative foodborne pathogenic bacteria.

The object of this study was to elucidate the bactericidal mechanism of a 222 nm Krypton Chlorine (KrCl) excilamp compared with that of a 254 nm Low Pressure mercury (LP Hg) lamp. The KrCl excilamp had higher bactericidal capacity against Gram-positive pathogenic bacteria (Staphylococcus aureus and L. monocytogenes) and Gram-negative pathogenic bacteria (S. Typhimurium and E. coli O157:H7) than did the LP Hg lamp when cell suspensions in PBS were irradiated with each type of UV lamp. It was found out that the KrCl excilamp induced cell membrane damage as a form of depolarization. From the study of respiratory chain dehydrogenase activity and the lipid peroxidation assay, it was revealed that cell membrane damage was attributed to inactivation of enzymes related to generation of membrane potential and occurrence of lipid peroxidation. Direct absorption of UV radiation which led to photoreaction through formation of an excited state was one of the causes inducing cell damage. Additionally, generation of ROS and thus occurrence of secondary damage can be another cause. The LP Hg lamp only induced damage to DNA but not to other components such as lipids or proteins. This difference was derived from differences of UV radiation absorption by cellular materials.

Effect of Laser Irradiation on Cell Function and Its Implications in Raman Spectroscopy.

Lasers are instrumental in advanced bioimaging and Raman spectroscopy. However, they are also well known for their destructive effects on living organisms, leading to concerns about the adverse effects of laser technologies. To implement Raman spectroscopy for cell analysis and manipulation, such as Raman-activated cell sorting, it is crucial to identify nondestructive conditions for living cells. Here, we evaluated quantitatively the effect of 532-nm laser irradiation on bacterial cell fate and growth at the single-cell level. Using a purpose-built microfluidic platform, we were able to quantify the growth characteristics, i.e., specific growth rates and lag times of individual cells, as well as the survival rate of a population in conjunction with Raman spectroscopy. Representative Gram-negative and Gram-positive species show similar trends in response to a laser irradiation dose. Laser irradiation could compromise the physiological function of cells, and the degree of destruction is both dose and strain dependent, ranging from reduced cell growth to a complete loss of cell metabolic activity and finally to physical disintegration. Gram-positive bacterial cells are more susceptible than Gram-negative bacterial strains to irradiation-induced damage. By directly correlating Raman acquisition with single-cell growth characteristics, we provide evidence of nondestructive characteristics of Raman spectroscopy on individual bacterial cells. However, while strong Raman signals can be obtained without causing cell death, the variety of responses from different strains and from individual cells justifies careful evaluation of Raman acquisition conditions if cell viability is critical.IMPORTANCE In Raman spectroscopy, the use of powerful monochromatic light in laser-based systems facilitates the detection of inherently weak signals. This allows environmentally and clinically relevant microorganisms to be measured at the single-cell level. The significance of being able to perform Raman measurement is that, unlike label-based fluorescence techniques, it provides a "fingerprint" that is specific to the identity and state of any (unlabeled) sample. Thus, it has emerged as a powerful method for studying living cells under physiological and environmental conditions. However, the laser's high power also has the potential to kill bacteria, which leads to concerns. The research presented here is a quantitative evaluation that provides a generic platform and methodology to evaluate the effects of laser irradiation on individual bacterial cells. Furthermore, it illustrates this by determining the conditions required to nondestructively measure the spectra of representative bacteria from several different groups.

Effects of gamma irradiation on the physico-chemical and biological properties of levofloxacin.

The aim of the present study was to examine the effect of gamma radiation on levofloxacin. Powder form of levofloxacin was subjected to different radiation doses (25, 50, 75, 100 and 125kGy) of Cobalt-60 source in a Gammacell-220 at a rate of 8.5 Gray/hr. The effect of radiation has been investigated with the aid of different spectroscopic techniques (UV-Vis, FT-IR), scanning electron microscopy (SEM), X-ray diffraction (XRD), and by antibacterial activities. UV data did not reveal significant changes in the structure of levofloxacin which is supported by scanning electron microscopy. However, X-rays diffraction shows a change in crystallinity of levofloxacin to an amorphous structure and this has been reflected on the morphology of this compound as indicated by SEM images. The antibacterial activities, on the other hand, reveal resistance of irradiated levofloxacin against bacteria, where some bacteria were highly affected by the irradiated drug. Similarly, FT-IR data show some changes in the functional groups principal absorption bands, in the IR spectrum, at frequencies 3286, 2846, 1716 and 1620 cm-1 for the O-H stretching band of quinolone, C-H stretching band, and C=O stretching band of carboxylic and pyridine. In addition, new peaks appeared which were not seen in the non-irradiated spectrum. In conclusion, some changes occurred in levofloxacin drug with the passage of radiation but the drug was chemically stable.

Evaluation of a shoe sole UVC device to reduce pathogen colonization on floors, surfaces and patients.

BACKGROUND: An ultraviolet C (UVC) decontamination device that delivers germicidal UVC radiation to the soles of shoes has become available recently. AIM: To demonstrate that shoe soles can be vectors for healthcare-associated infection, and to investigate if a UVC shoe sole decontamination device would decrease this risk effectively. METHODOLOGY: Three bacterial strains (Staphylococcus aureus, Enterococcus faecalis and Escherichia coli) and a non-toxigenic strain of Clostridium difficile were spiked on to standardized rubber-soled shoe soles and then selected at random for UVC exposure or no UVC exposure. Experiments were performed to test the efficacy of the UVC device to decontaminate shoe soles and flooring. E. faecalis was spiked on to shoes to assess colonization of a simulated healthcare environment and patient. RESULTS: The UVC device decreased shoe sole contamination significantly for all tested bacterial species, and decreased floor contamination significantly for all floor types and species tested (P<0.01 for all experiments). The log10 reduction was the highest for E. coli (mean±standard deviation 2.6±0.79), followed by E. faecalis (2.19±0.68), S. aureus (1.74±0.88) and C. difficile (0.42±0.54) (P<0.0001 for all analyses). Exposure of shoe soles to the UVC device decreased contamination significantly (mean log10 reduction 2.79±1.25; P<0.0001). Proportions of samples from furniture, bed and patient dummy samples decreased from 96-100% positive in controls to 5-8% positive in UVC device experiments (P<0.0001 for all analyses). CONCLUSION: A UVC decontamination device was shown to reduce the colony-forming unit counts of relevant pathogenic organisms from shoe soles with subsequent decreased colonization of floors, healthcare equipment, furniture, beds and a patient dummy.

Photo-induced toxicity of tungsten oxide photochromic nanoparticles.

We synthesised a new type of photochromic tungsten oxide nanoparticles, analysed their photocatalytic activity and carried out a thorough analysis of their effect on prokaryotic and eukaryotic organisms. Ultrasmall hydrated tungsten oxide nanoparticles were prepared by means of hydrothermal treatment of tungstic acid in the presence of polyvinylpyrrolidone as a template, stabiliser and growth regulator. Tungstic acid was synthesised through an ion-exchange method using sodium tungstate solution and a strongly acidic cation exchange resin. Upon illumination, photochromic nanoparticles of WO3 were shown to increase greatly their toxicity against both bacterial (both gram-positive and gram-negative - P. aeruginosa, E. coli and S. aureus) and mammalian cells (primary mouse embryonic fibroblasts); under the same conditions, fungi (C. albicans) were less sensitive to the action of tungsten oxide nanoparticles. UV irradiation of primary mouse fibroblasts in the presence of WO3 nanoparticles demonstrated a time- and dose-dependent toxic effect, the latter leading to a significant decrease in dehydrogenase activity and an increase in the number of dead cells. WO3 nanoparticles were photocatalytically active under both UV light and even diffused daylight filtered through a window glass, leading to indigo carmine organic dye discolouration. The obtained experimental data not only show good prospects for biomedical applications of tungsten trioxide, but also demonstrate the need for clear control of biosafety when it is used in various household materials and appliances.