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Background: Acinetobacter baumannii , a nosocomial pathogen, poses a major public health problem due to generating resistance to several antimicrobial agents. Antimicrobial photodynamic inactivation (APDI) employs a nontoxic dye as a photosensitizer (PS) and light to produce reactive oxygen species that destroy bacterial cells. The intracellular concentration of PS could be affected by factors such as the function of efflux pumps to emit PS from the cytosol. Objective: To evaluate the augmentation effect of an efflux pump inhibitor, verapamil, three multidrug-resistant A. baumannii were subjected to APDI by erythrosine B (EB). Methods and results: The combination of EB and verapamil along with irradiation at 530 nm induced a lethal effect and more than 3 log colony-forming unit reduction to all A. baumannii strains in planktonic state. In contrast, EB and irradiation alone could produce only a sublethal effect on two of the strains. Conclusions: These data suggest that verapamil increases the intracellular concentration of EB, which potentiates the lethal efficacy of APDI. Verapamil could be applied with EB and green light to improve their antimicrobial efficacy against A. baumannii -localized infections.

The spread of antibiotic resistant pathogens and antibiotic resistance genes (ARGs) in the environment poses a serious threat to public health. However, existing methods are difficult to effectively remove antibiotic resistant pathogens and ARGs from the environment. In this study, we synthesized a new acridine-based photosensitizer, 2,7-dibromo-9-mesityl-10-methylacridinium perchlorate (YM-3), by the heavy atom effect, which could photodynamically inactivate antibiotic resistant pathogens and reduce ARGs by generating singlet oxygen ( 1 O 2 ) in an aqueous environment. The 1 O 2 yield of YM-3 was 4.9 times that of its modified precursor. YM-3 could reduce the culturable number and even the viable counts of methicillin-resistant Staphylococcus aureus and carbapenem-resistant Acinetobacter baumannii to 0 (inactivation rate > 99.99999%) after 2 and 8 h of low-intensity blue light (15 W/m 2 ) irradiation, respectively. After 20 h of light exposure, the copy numbers of ARGs in both bacteria were reduced by 5.80 and 4.48 log, respectively, which might indicate that ARGs had been degraded. In addition, YM-3 still had an efficient bactericidal effect after five inactivation cycle. These characteristics of ultra-low light intensity requirement and efficient bactericidal ability make YM-3 have good application prospects for disinfection in indoor and sunlight environment., Competing Interests: Declaration of Competing Interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2023 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Many considerable investigations focused on the stimulation of therapeutic manners of infected injuries in mice. The exaggerated pathogens that induced wounds were gram-positive like staphylococcal and gram-negative, for example, Pseudomonas aeuroginosa and Acinetobacter baumannii . Acinetobacter can generate a scale range of an infection that may be received in a hospital or any wellness concern facility. In order to know the significance of laser 532 nm with a constant irradiance at various exposure times on the healing process of wounds infected by Acinetobacter baumannii , this study was performed on the BALB/C mice. An elliptical full-thickness skin injury was made on the backside of 45 adult female (BALB/C) mice. Injuries were affected via Acinetobacter baumannii and were randomly assigned into 3 groups. Semiconductor diode continuous wave laser, λ = 532 nm, with output power 40 mW was used. The power density was 5.71 mW/cm 2 , while the fluencies were 1.7 J/cm 2 and 5.14 J/cm 2 . Fifteen mice were classified according to the times of irradiation. The first group was infected and presented as control, without irradiation. The second group was infected and irradiated for 5 minutes. The third group, likewise, was infected but irradiated for 15 minutes. All groups were subdivided according to the following period, 3, 5, and 10 days, after irradiation and the animals were killed after the treatment. Wound healing was made by measuring the rate of wound closure and histopathological evaluation. The study determined that 532 nm laser therapy had an obvious and positive influence on the healing of infected wounds with fluence (5.14 J/cm 2 ).

Light sensing has been extensively characterized in the human pathogen Acinetobacter baumannii at environmental temperatures. However, the influence of light on the physiology and pathogenicity of human bacterial pathogens at temperatures found in warm-blooded hosts is still poorly understand. In this work, we show that Staphylococcus aureus , Acinetobacter baumannii , and Pseudomonas aeruginosa (ESKAPE) priority pathogens, which have been recognized by the WHO and the CDC as critical, can also sense and respond to light at temperatures found in human hosts. Most interestingly, in these pathogens, light modulates important pathogenicity determinants as well as virulence in an epithelial infection model, which could have implications in human infections. In fact, we found that alpha-toxin-dependent hemolysis, motility, and growth under iron-deprived conditions are modulated by light in S. aureus Light also regulates persistence, metabolism, and the ability to kill competitors in some of these microorganisms. Finally, light exerts a profound effect on the virulence of these pathogens in an epithelial infection model, although the response is not the same in the different species; virulence was enhanced by light in A. baumannii and S. aureus , while in A. nosocomialis and P. aeruginosa it was reduced. Neither the BlsA photoreceptor nor the type VI secretion system (T6SS) is involved in virulence modulation by light in A. baumannii Overall, this fundamental knowledge highlights the potential use of light to control pathogen virulence, either directly or by manipulating the light regulatory switch toward the lowest virulence/persistence configuration. IMPORTANCE Pathogenic bacteria are microorganisms capable of producing disease. Dangerous bacterial pathogens, such as Staphylococcus aureus , Pseudomonas aeruginosa , and Acinetobacter baumannii , are responsible for serious intrahospital and community infections in humans. Therapeutics is often complicated due to resistance to multiple antibiotics, rendering them ineffective. In this work, we show that these pathogens sense natural light and respond to it by modulating aspects related to their ability to cause disease; in the presence of light, some of them become more aggressive, while others show an opposite response. Overall, we provide new understanding on the behavior of these pathogens, which could contribute to the control of infections caused by them. Since the response is distributed in diverse pathogens, this notion could prove a general concept., (Copyright © 2021 American Society for Microbiology.)

Background and Objectives: Biofilms cause more than 80% of infections in humans, including more than 90% of all chronic wound infections and are extremely resistant to antimicrobials and the immune system. The situation is exacerbated by the fast spreading of antimicrobial resistance, which has become one of the biggest threats to current public health. There is consequently a critical need for the development of alternative therapeutics. Antimicrobial blue light (aBL) is a light-based approach that exhibits intrinsic antimicrobial effect without the involvement of exogenous photosensitizers. In this study, we investigated the antimicrobial effect of this non-antibiotic approach against biofilms formed by microbial isolates of multidrug-resistant bacteria., Study Design/materials and Methods: Microbial isolates of Acinetobacter baumannii, Candida albicans, Escherichia coli, Enterococcus faecalis, MRSA, Neisseria gonorrhoeae, Pseudomonas aeruginosa, and Proteus mirabilis were studied. Biofilms were grown in microtiter plates for 24 or 48 hours or in the CDC biofilm reactor for 48 hours and exposed to aBL at 405 nm (60 mW/cm 2 , 60 or 30 minutes). The anti-biofilm activity of aBL was measured by viable counts., Results: The biofilms of A. baumannii, N. gonorrhoeae, and P. aeruginosa were the most susceptible to aBL with between 4 and 8 log 10 inactivation after 108 J/cm 2 (60 mW/cm 2 , 30 minutes) or 216 J/cm 2 (60 mW/cm 2 , 60 minutes) aBL were delivered in the microplates. On the contrary, the biofilms of C. albicans, E. coli, E. faecalis, and P. mirabilis were the least susceptible to aBL inactivation (-0.30, -0.24, -0.84, and -0.68 log 10 inactivation, respectively). The same aBL treatment in biofilms developed in the CDC biofilm reactor, caused -1.68 log 10 inactivation in A. baumannii and -1.74 and -1.65 log 10 inactivation in two different strains of P. aeruginosa., Conclusions: aBL exhibits potential against pathogenic microorganisms and could help with the significant need for new antimicrobials in clinical practice to manage multidrug-resistant infections. Lasers Surg. Med. © 2019 Wiley Periodicals, Inc., (© 2019 Wiley Periodicals, Inc.)

Rationale: Antimicrobial photodynamic treatment is potentially an alternative to antibiotics and is also effective against viruses, fungi and some cancers. Our previous studies have shown that blue light combined with curcumin, a chemical from the turmeric plant, exerted effective antimicrobial activity via photodynamic treatment. The study reported in this paper investigates which target proteins are affected after the treatment., Methods: We treated imipenem-resistant Acinetobacter baumannii with blue light and curcumin and used protein carbonylation as a marker for oxidative damage. After treatment, the bacterial proteins were extracted and the protein carbonyls marked using dinitrophenylhydrazide. After enzyme digestion, we used liquid chromatography/nano-electrospray ionization (LC/nano-ESI) ion trap mass spectrometry to identify bacterial peptides from a customized database. The functional enrichment analyses of the identified proteins were performed using gene ontology annotation and the STRING protein-protein interaction network., Results: The application of curcumin with blue light showed good antibacterial activity against imipenem-resistant A. baumannii. Using a shotgun proteomics approach, the carbonylated proteins in A. baumannii caused by the photolytic curcumin were identified. The results showed that the proteins related to membrane structures, translation and response to oxidative stress were preferentially modified., Conclusions: The photolytic curcumin treatment could be a potential alternative to antibiotics for bacterial infection. In this study, the shotgun proteomics strategy allows us to explore the possible bactericidal mechanisms under this oxidative stress. The result provides a reference for future studies on the enhancement of the action of photolytic curcumin., (© 2019 John Wiley & Sons, Ltd.)

Background: Ultraviolet (UV) fluorescent lamp (FL) was applied in mainstream riboflavin photochemical method (RPM) to inactivate pathogens in blood components. Low UV irradiance emitted by UV-FL resulted in more time to achieve effective inactivation., Materials and Methods: A novel light emitting diode (LED) UV illumination with adjustable irradiance was developed by us. Two strains of drug-resistant bacteria (DRB), pan-drug resistant Acinetobacter baumannii (PDRAB) and methicillin-resistant Staphylococcus aureus (MRSA) were cultured and used for evaluating the inactivation effectiveness of RPM using UV-LED or UV-FL against DRB in plasma or platelets. Three plasma factors and four platelet parameters were measured after treatments., Results: There was a linear relationship between UV-LED irradiance and electric current, the minimum UV irradiance was 24 mW/cm 2 , and the maximum was 258 mW/cm 2 . At the same UV dose of 15 J/cm 2 , inactivation effectiveness of UV-LED with 258 mW/cm 2 against PDRAB in plasma or platelets were comparable to that of UV-FL with 16 mW/cm 2 , both above 98%. UV-FL treatment required 10-15 min, but UV-LED only required 1-2 min. However, MRSA showed a resistance to UV-LED (inactivation effectiveness was around 40%) compared with UV-FL (inactivation effectiveness was above 98%). The retention of fibrinogen, factor V, factor VII in plasma and platelet counts in platelets with UV-LED treatment were significantly higher than UV-FL at the same UV dose., Conclusion: The treatment of RPM using UV-LED with high UV irradiance was able to dramatically shorten inactivation time against PDRAB in plasma or platelets and improve retention of blood components compared with UV-FL., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Background: Antimicrobial resistance is a significant concern to public health, and there is a pressing need to develop novel antimicrobial therapeutic modalities., Methods: In this study, we investigated the capacity for quinine hydrochloride (Q-HCL) to enhance the antimicrobial effects of antimicrobial blue light ([aBL] 405 nm wavelength) against multidrug-resistant (MDR) Gram-negative bacteria in vitro and in vivo., Results: Our findings demonstrated the significant improvement in the inactivation of MDR Pseudomonas aeruginosa and Acinetobacter baumannii (planktonic cells and biofilms) when aBL was illuminated during Q-HCL exposure. Furthermore, the addition of Q-HCL significantly potentiated the antimicrobial effects of aBL in a mouse skin abrasion infection model. In addition, combined exposure of aBL and Q-HCL did not result in any significant apoptosis when exposed to uninfected mouse skin., Conclusions: In conclusion, aBL in combination with Q-HCL may offer a novel approach for the treatment of infections caused by MDR bacteria., (© The Author(s) 2019. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail: journals.permissions@oup.com.)

Background: No-touch environmental disinfection using ultraviolet devices has been highlighted in the past several years to control the transmission of multidrug-resistant organisms (MDROs). However, its effectiveness in non-US healthcare settings is yet to be examined. This study aimed to evaluate the effectiveness of disinfection by portable pulsed xenon ultraviolet (PX-UV) devices in controlling transmission of MDROs in a non-US healthcare setting., Methods: All patients admitted in the intensive care unit in a 629-bed tertiary referral hospital in Japan from August 2016 to February 2019 were enrolled. During the study period, PX-UV disinfection was added to manual terminal cleaning after every patient transfer/discharge. For microbiological evaluation, surfaces were selected for sampling by contact plates before/after manual cleaning and after PX-UV. After overnight incubation, colonies on the plates were counted., Results: The incidence of newly acquired methicillin-resistant Staphylococcus aureus (MRSA) declined significantly (13.8 to 9.9 per 10,000 patient days, incidence rate ratio 0.71, p = 0.002), as well as that of newly acquired drug-resistant Acinetobacter (48.5 to 18.1, 0.37, p < 0.001). The percent reduction of the microbiological burden by manual cleaning was 81%, but a further 59% reduction was achieved by PX-UV., Conclusions: PX-UV is effective in further reducing the microbial burden and controlling MDROs in a non-US healthcare setting.

Light-emitting diodes (LEDs) demonstrate therapeutic effects for a range of biomedical applications, including photodisinfection. Bands of specific wavelengths (centered at 405 nm) are reported to be the most antimicrobial; however, there remains no consensus on the most effective irradiation parameters for optimal photodisinfection. The aim of this study was to assess decontamination efficiency by direct photodisinfection of monomicrobial biofilms using a violet-blue light (VBL) single-wavelength array (SWA) and multiwavelength array (MWA). Mature biofilms of nosocomial bacteria ( Acinetobacter baumannii , Pseudomonas aeruginosa , Escherichia coli , and Staphylococcus aureus ) were grown on 96-well polypropylene PCR plates. The biofilms were then exposed to VBL for 2,700 s (SWA) and 1,170 s (MWA) to deliver 0 to 670 J/cm 2 , and the antibacterial activity of VBL was assessed by comparing the seeding of the irradiated and the nonirradiated biofilms. Nonirradiated groups were used as controls. The VBL arrays were characterized optically (spectral irradiance and beam profile) and thermally. The SWA delivered 401-nm VBL and the MWA delivered between 379-nm and 452-nm VBL, albeit at different irradiances and with different beam profiles. In both arrays, the irradiated groups were exposed to increased temperatures compared to the nonirradiated controls. All bacterial isolates were susceptible to VBL and demonstrated reductions in the seeding of exposed biofilms compared with the nonirradiated controls. VBL at 405 nm exerted the most antimicrobial activity, exhibiting reductions in seeding of up to 94%. Decontamination efficiency is dependent on the irradiation parameters, bacterial species and strain, and experimental conditions. Controlled experiments that ameliorate the heating effects and improve the optical properties are required to optimize the dosing parameters to advance the successful clinical translation of this technology. IMPORTANCE This study reports the efficacy of VBL and blue light (BL) and their antimicrobial activity against mature biofilms of a range of important nosocomial pathogens. While this study investigated the antibacterial activity of a range of wavelengths of between 375 and 450 nm and identified a specific wavelength region (∼405 nm) with increased antibacterial activity, decontamination was dependent on the bacterial species, strain, irradiation parameters, and experimental conditions. Further research with controlled experiments that ameliorate the heating effects and improve the optical properties are required to optimize the dosing parameters to advance the successful clinical translation of this technology., (Copyright © 2019 Halstead et al.)

Sunlight is a ubiquitous environmental stimulus for the great majority of living organisms on Earth; therefore it is logical to expect the development of "seeing mechanisms" which lead them to successfully adapt to particular ecological niches. Although these mechanisms were recognized in photosynthetic organisms, it was not until recent years that the scientific community found out about light perception in chemotrophic ones. In this review we summarize the current knowledge about the mechanism of light sensing through the blue light receptor BlsA in Acinetobacter baumannii. We highlight its function as a global regulator that pleiotropically modulates a large number of physiological processes, many of which are linked to the ability of this opportunist pathogen to persist in adverse intrahospital environments. Moreover, we describe with some specific examples the molecular basis of how this photoregulator senses blue light and translates this physical signal by modulating gene expression of target regulons. Finally, we discuss the possible course of these investigations needed to dissect this complex regulatory network, which ultimately will help us better understand the A. baumannii physiology.

Background: Acinetobacter baumannii is a cause of healthcare-associated infections and has considerable potential to survive on inanimate hospital surfaces under hostile conditions (e.g. disinfection or desiccation)., Aim: To learn more about its survival strategy and capacity to persist in liquid media and on surfaces mimicking hospital environments., Methods: The effect of temperature, nutrient deprivation, permanence on inanimate surfaces, and exposure to disinfectants on the survival of four A. baumannii strains (ATCC 19606 T and three clinical isolates) was studied by monitoring the number of total and viable cells using fluorescent microscopy and of culturable cells by standard cultures., Findings: Bacterial survival was differentially affected by temperature (cells maintained at 20°C remained culturable at least within 30 days) and physical environment (desiccation favoured cell resistance to stress at 37°C). Moreover, persistence was associated with two adaptation patterns: one linked to entry into the viable but non-culturable state, whereas the other apparently followed a bust-and-boom model. During a study on the effect of disinfectant (commercial bleach and quaternary ammonium compounds), it was found that treatment with these antibacterial compounds did not eliminate A. baumannii populations and provoked the reduction of culturable populations, although a fraction of cells remained culturable., Conclusion: The ability to persist for long periods on different surfaces, mimicking those usually found in hospitals, along with A. baumannii's capacity to survive after a disinfection process may account for the recurrent outbreaks in intensive care units., (Copyright © 2019 The Healthcare Infection Society. Published by Elsevier Ltd. All rights reserved.)

Background: Healthcare-associated infections (HAIs) caused by multidrug-resistant bacteria (MDRB) are of global concern and hospital textiles can contribute to their transmission. MDRB are able to survive on textiles for more than enough time to spread in the environment. Some studies summarized the effect of environmental factors on the duration of bacterial survival, but it remained an open question how these factors influence the quantity of surviving bacteria in a period of a few days, which is relevant from the perspective of HAIs. Investigating this effect can contribute to better understand the spread of MDRB and the emergence of hospital outbreaks., Methods: We investigated quantitatively the survival capability of 15 vancomycin-resistant Enterococcus faecium (VRE), 15 methicillin-resistant Staphylococcus aureus (MRSA), 15 multidrug-resistant Acinetobacter baumannii (MACI) and 15 multidrug-resistant Klebsiella pneumoniae (MRKP) in five environmental conditions using the plate count method. We examined the role of nutrients, textile types, temperature and level of relative humidity on bacterial survival after 1-7days of incubation., Results: Each bacterial group showed higher survival capability on 100% cotton towel than on 100% cotton sheet (P<0.01). MRSAs and VREs showed higher (P<0.01), MACIs showed lower (P=0.02) CFU/swatch values on 100% polyester sheet than on cotton sheet. The survival capability of MRKPs and MRSAs was higher inoculated in nutrient broth than in saline solution (P<0.01). Each bacterial group showed lower survival capability (P<0.01) at body condition (T=35°C, Rh=83%) than at control (T=25°C, Rh=52%)., Conclusions: Towels proved to be excellent conditions for each bacteria to survive, however chemical composition of the textiles affected differently the survival of Gram-positive and Gram-negative bacteria. These findings could be useful in searching for the source of outbreaks. Organic contamination of the textiles can increase the survival of desiccation-sensitive bacteria, therefore nutrient-rich inoculating medium is recommended in survival studies., (Copyright © 2018 The Authors. Published by Elsevier Ltd.. All rights reserved.)

Light modulates global features of the important human pathogen Acinetobacter baumannii lifestyle including metabolism, tolerance to antibiotics and virulence, most of which depend on the short BLUF-type photoreceptor BlsA. In this work, we show that the ability to circumvent iron deficiency is also modulated by light at moderate temperatures, and disclose the mechanism of signal transduction by showing that BlsA antagonizes the functioning of the ferric uptake regulator (Fur) in a temperature-dependent manner. In fact, we show that BlsA interacts with Fur in the dark at 23 °C, while the interaction is significantly weakened under blue light. Moreover, under iron deprived conditions, expression of Fur-regulated Acinetobactin siderophore genes is only induced in the dark in a BlsA-dependent manner. Finally, growth under iron deficiency is supported in the dark rather than under blue light at moderate temperatures through BlsA. The data is consistent with a model in which BlsA might sequester the repressor from the corresponding operator-promoters, allowing Acinetobactin gene expression. The photoregulation of iron metabolism is lost at higher temperatures such as 30 °C, consistent with fading of the BlsA-Fur interaction at this condition. Overall, we provide new understanding on the functioning of the widespread Fur regulator as well as short-BLUFs.

Photodynamic inactivation (PDI) is a non-invasive and safe therapeutic method for microbial infections. Bacterial antibiotic resistance is caused by antibiotics abuse. Drug-resistant Acinetobacter spp. is a serious problem in hospitals around the world. These pathogens from nosocomial infections have high mortality rates in frailer people, and Acinetobacter spp. is commonly found in immunocompromised patients. Visible light is safer than ultraviolet light (UV) for PDI of nosocomial pathogens with mammalian cells. Zinc oxide nanoparticles (ZnO-NPs) were used in this study as an antimicrobial agent and a photosensitizer. ZnO is recognized as safe and has extensive usage in food additives, medical and cosmetic products. In this study, we used 0.125 mg/ml ZnO-NPs combined with 10.8 J/cm 2 blue light (BL) on Acinetobacter baumannii (A. baumannii) that could significantly reduce microbial survival. However, individual exposure to ZnO-NPs does not affect the viability of A. baumannii. BL irradiation could trigger the antimicrobial ability of ZnO nanoparticles on A. baumannii. The mechanism of photocatalytic ZnO-NPs treatment for sterilization occurs through bacterial membrane disruptions. Otherwise, the photocatalytic ZnO-NPs treatment showed high microbial eradication in nosocomial pathogens, including colistin-resistant and imipenem-resistant A. baumannii and Klebsiella pneumoniae. Based on our results, the photocatalytic ZnO-NPs treatment could support hygiene control and clinical therapies without antibiotics to nosocomial bacterial infections., (Copyright © 2018. Published by Elsevier B.V.)

Nosocomially-acquired multi-, extensively-, and pandrug resistant (MDR, XDR, and PDR) strains of microorganisms such as Acinetobacter baumannii remain a serious cause of infection and septic mortality in burn patients. Treatment of patients with nosocomial burn wound infections is often complicated by drug-resistant strains of A. baumannii. Today, many researchers are focusing on the investigation of novel non-antibiotic strategies such as photodynamic therapy (PDT). We report a new PDT strategy that suppresses colistin resistance in PDR A. baumannii by interfering with the expression of a pmrA/pmrB two-component system. In the current study, A. baumannii with a PDR feature isolated from a burn patient was used as a test strain. PDT was carried out using toluidine blue O (TBO) and light-emitting diode (LED) as a photosensitizer and radiation source, respectively. The antimicrobial susceptibility profiles were assessed for cells surviving PDT. The effects of sub-lethal PDT (sPDT) on the expression of the pmrA/pmrB two-component signal transduction system were evaluated by real-time quantitative reverse transcription PCR. Results of drug susceptibly testing (DST) in LED and TBO groups separately showed that the bacteria were resistant to all tested antibiotics, while the DST result of the LED+TBO group showed highly declining bacterial growth when compared with the control group. Reduction in the expression of pmrA and pmrB was observed in the treated strains after sPDT. This represents the first conclusive example of a direct role for the PDT in breaking antibiotic resistance by directly modulating two-component system activity., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Photodynamic antimicrobial chemotherapy (PACT), as a novel and effective therapeutic modality to eradicate drug resistant bacteria without provoking multidrug resistance, has attracted increasing attention. This study examined the antimicrobial efficacy of the novel cationic amino acid-porphyrin conjugate 4I with four lysine groups against two different clinical isolated strains (drug sensitive and multidrug resistant) of the Acinetobacter baumannii species and its toxicity on murine dermal fibroblasts in vitro, as well as the therapeutic effect of PACT on acute, potentially lethal multidrug resistant strain excisional wound infections in vivo. The PACT protocol exposed 4I to illumination, exhibiting high antimicrobial efficacy on two different strains due to a high yield of reactive oxygen species (ROS) and non-selectivity to microorganisms. The photoinactivation effects of 4I against two different strains were dose-dependent. At 3.9 μM and 7.8 μM, PACT induced 6 log units of inactivation of sensitive and multidrug resistant strains. In contrast, 4I alone and illumination alone treatments had no visibly antimicrobial effect. Moreover, cytotoxicity tests revealed the great safety of the photosensitizer 4I in mice. In the in vivo study, we found 4I-mediated PACT was not only able to kill bacteria but also accelerated wound recovery. Compared with non-treated mice, over 2.89 log reduction of multidrug resistant Acinetobacter baumannii strain was reached in PACT treat mice at 24 h post-treatment. These results imply that 4I-mediated PACT therapy is an effective and safe alternative to conventional antibiotic therapy and has clinical potential for superficial drug-resistant bacterial infections.

In the nosocomial opportunistic pathogen Acinetobacter baumannii , RecA-dependent mutagenesis, which causes antibiotic resistance acquisition, is linked to the DNA damage response (DDR). Notably, unlike the Escherichia coli paradigm, recA and DDR gene expression in A. baumannii is bimodal. Namely, there is phenotypic variation upon DNA damage, which may provide a bet-hedging strategy for survival. Thus, understanding recA gene regulation is key to elucidate the yet unknown DDR regulation in A. baumannii Here, we identify a structured 5' untranslated region (UTR) in the recA transcript which serves as a cis -regulatory element. We show that a predicted stem-loop structure in this 5' UTR affects mRNA half-life and underlies bimodal gene expression and thus phenotypic variation in response to ciprofloxacin treatment. We furthermore show that the stem-loop structure of the recA 5' UTR influences intracellular RecA protein levels and, in vivo , impairing the formation of the stem-loop structure of the recA 5' UTR lowers cell survival of UV treatment and decreases rifampin resistance acquisition from DNA damage-induced mutagenesis. We hypothesize that the 5' UTR allows for stable recA transcripts during stress, including antibiotic treatment, enabling cells to maintain suitable RecA levels for survival. This innovative strategy to regulate the DDR in A. baumannii may contribute to its success as a pathogen. IMPORTANCE Acinetobacter baumannii is an opportunistic pathogen quickly gaining antibiotic resistances. Mutagenesis and antibiotic resistance acquisition are linked to the DNA damage response (DDR). However, how the DDR is regulated in A. baumannii remains unknown, since unlike most bacteria, A. baumannii does not follow the regulation of the Escherichia coli paradigm. In this study, we have started to uncover the mechanisms regulating the novel A. baumannii DDR. We have found that a cis -acting 5' UTR regulates recA transcript stability, RecA protein levels, and DNA damage-induced phenotypic variation. Though 5' UTRs are known to provide stability to transcripts in bacteria, this is the first example in which it regulates a bimodal DDR response through recA transcript stabilization, potentially enabling cells to have enough RecA for survival and genetic variability., (Copyright © 2017 American Society for Microbiology.)

Light sensing in chemotrophic bacteria has been relatively recently ascertained. In the human pathogen Acinetobacter baumannii , light modulates motility, biofilm formation, and virulence through the blue-light-sensing-using flavin (BLUF) photoreceptor BlsA. In addition, light can induce a reduction in susceptibility to certain antibiotics, such as minocycline and tigecycline, in a photoreceptor-independent manner. In this work, we identified new traits whose expression levels are modulated by light in this pathogen, which comprise not only important determinants related to pathogenicity and antibiotic resistance but also metabolic pathways, which represents a novel concept for chemotrophic bacteria. Indeed, the phenylacetic acid catabolic pathway and trehalose biosynthesis were modulated by light, responses that completely depend on BlsA. We further show that tolerance to some antibiotics and modulation of antioxidant enzyme levels are also influenced by light, likely contributing to bacterial persistence in adverse environments. Also, we present evidence indicating that surfactant production is modulated by light. Finally, the expression of whole pathways and gene clusters, such as genes involved in lipid metabolism and genes encoding components of the type VI secretion system, as well as efflux pumps related to antibiotic resistance, was differentially induced by light. Overall, our results indicate that light modulates global features of the A. baumannii lifestyle. IMPORTANCE The discovery that nonphototrophic bacteria respond to light constituted a novel concept in microbiology. In this context, we demonstrated that light could modulate aspects related to bacterial virulence, persistence, and resistance to antibiotics in the human pathogen Acinetobacter baumannii In this work, we present the novel finding that light directly regulates metabolism in this chemotrophic bacterium. Insights into the mechanism show the involvement of the photoreceptor BlsA. In addition, tolerance to antibiotics and catalase levels are also influenced by light, likely contributing to bacterial persistence in adverse environments, as is the expression of the type VI secretion system and efflux pumps. Overall, a profound influence of light on the lifestyle of A. baumannii is suggested to occur., (Copyright © 2017 American Society for Microbiology.)

Objectives: To determine the impact of long-term use of antibiotics and ultraviolet radiation on the resistance of Acinetobacter baumannii to tigecycline and the viability of tigecycline-resistant Acinetobacter baumannii ., Methods: Three strains of tigecycline sensitive Acinetobacter baumannii were selected. Tigecycline resistance was induced through multi-step method or by ultraviolet radiation. Two strains of tigecycline resistant Acinetobacter baumannii were repeatedly passaged on blank MHA plates, for the purpose of determination of minimum inhibitory concentrations (MICs) of tigecycline using broth microdilution method. The tigecycline sensitive (b38) and homologous resistant Acinetobacter baumannii (b38!d) were cultured separately and conjointly to evaluate its fitness costs of tigecycline resistance., Results: Tigecycline resistant strains were successfully induced using multi-step method. Ultraviolet radiation did not change the sensitivity of the three strains to tigecycline, but elevated the MICs of tigecycline. The MICs of tigecycline did not change over 40 generations. It took much more time for the resistant strains to reach logarithmic growth phase and plateau phase compared with the tigecycline sensitive strains. With repeated passage, the tigecycline resistant strains decreased rapidly, even vanished in conjointly culture., Conclusion: Acinetobacter baumannii can acquire tigecycline resistance. The resistance may have genetic stability. The resistant strains have less adaptability than the sensitive strains.

Introduction: Widespread resistance to antimicrobial agents has led to a dearth of therapeutic choices in treating Acinetobacter baumannii infections, leading to new strategies for treatment being needed. We evaluated the effects of photodynamic therapy (PDT) as an alternative antimicrobial modality on the virulence features of cell-surviving PDT., Materials and Methods: To determine the sublethal PDT (sPDT), a colistin-resistant, extensively drug-resistant A. baumannii (CR-XDR-AB) clinical isolate and A. baumannii and ATCC 19606 strains, photosensitized with toluidine blue O (TBO), were irradiated with light emitting diodes, following bacterial viability measurements. The biofilm formation ability, outer membrane (OM) integrity, and antimicrobial susceptibility profiles were assessed for cell-surviving PDT. The effects of sPDT on the expression of virulent genes were evaluated by real-time quantitative reverse transcription PCR (qRT-PCR)., Results: sPDT resulted in the reduction of the biofilm formation capacity, and its metabolic activity in strains. The OM permeability and efflux pump inhibition of the sPDT-treated CR-XDR-AB cells were increased; however, there was no significant change in OM integrity in ATCC 19606 strain after sPDT. sPDT reduced the minimum inhibitory concentrations of the most tested antimicrobials by ≥2-fold in CR-XDR-AB. lpsB, blsA, and dnaK were upregulated after the strains were treated with sPDT; however, a reduction in the expression of csuE, epsA, and abaI was observed in the treated strains after sPDT., Conclusion: The susceptibility of CR-XDR-AB to a range of antibiotics was enhanced following sPDT. The virulence of strains is reduced in cells surviving PDT with TBO, and this may have potential implications of PDT for the treatment of A. baumannii infections., (Copyright © 2016 Elsevier B.V. All rights reserved.)

Background: Biofilms affect >80% bacterial infections in human and are usually difficult to eradicate because of their inherent drug resistance., Methods: We investigated the effectiveness of antimicrobial blue light (aBL) (wavelength, 415 nm) for inactivating Acinetobacter baumannii or Pseudomonas aeruginosa biofilms in 96-well microplates or infected mouse burn wounds., Results: In vitro, in 96-well microplates, exposure of 24-hour-old and 72-hour-old A. baumannii biofilms to 432 J/cm(2) aBL resulted in inactivation of 3.59 log10 and 3.18 log10 colony-forming units (CFU), respectively. For P. aeruginosa biofilms, similar levels of inactivation-3.02 log10 and 3.12 log10 CFU, respectively-were achieved. In mouse burn wounds infected with 5 × 10(6) CFU ofA. baumannii, approximately 360 J/cm(2) and 540 J/cm(2) aBL was required to inactivate 3 log10 CFU in biofilms when delivered 24 and 48 hours, respectively, after bacterial inoculation. High-performance liquid chromatography analysis revealed the presence of endogenous porphyrins in both A. baumannii and P. aeruginosa TUNEL assay detected no apoptotic cells in aBL-irradiated mouse skin at up to 24 hours after aBL exposure (540 J/cm(2))., Conclusions: aBL has antimicrobial activity in biofilms ofA. baumannii and P. aeruginosa and is a potential therapeutic approach for biofilm-related infections., (© The Author 2016. Published by Oxford University Press for the Infectious Diseases Society of America. All rights reserved. For permissions, e-mail journals.permissions@oup.com.)

Minocycline (MIN) and tigecycline (TIG) are antibiotics currently used for treatment of multidrug-resistant nosocomial pathogens. In this work, we show that blue light, as well as white light, modulates susceptibility to these antibiotics in a temperature-dependent manner. The modulation of susceptibility by light depends on the content of iron; an increase in iron results in a reduction in antibiotic susceptibility both under light and in the dark, though the effect is more pronounced in the latter condition. We further provide insights into the mechanism by showing that reduction in susceptibility to MIN and TIG induced by light is likely triggered by the generation of (1)O2, which, by a yet unknown mechanism, would ultimately lead to the activation of resistance genes such as those coding for the efflux pump AdeABC. The clinical relevance of these results may lie in surface-exposed wound infections, given the exposure to light in addition to the relatively low temperatures recorded in this type of lesion. We further show that the modulation of antibiotic susceptibility occurs not only in Acinetobacter baumannii but also in other micro-organisms of clinical relevance such as Escherichia coli and Staphylococcus aureus. Overall, our findings allow us to suggest that MIN and TIG antibiotic treatments may be improved by the inclusion of an iron chelator, in addition to keeping the wounds in the dark, a condition that would increase the effectiveness in the control of infections involving these micro-organisms., (© 2015 The Authors.)

Photodynamic inactivation of bacteria (PIB) proves to be an additional method to kill pathogenic bacteria. PIB requires photosensitizer molecules that effectively generate reactive oxygen species like singlet oxygen when exposed to visible light. To allow a broad application in medicine, photosensitizers should be safe when applied in humans. Substances like vitamin B2, which are most likely safe, are known to produce singlet oxygen upon irradiation. In the present study, we added positive charges to flavin derivatives to enable attachment of these molecules to the negatively charged surface of bacteria. Two of the synthesized flavin derivatives showed a high quantum yield of singlet oxygen of approximately 75%. Multidrug resistant bacteria like MRSA (Methicillin resistant Staphylococcus aureus), EHEC (enterohemorrhagic Escherichia coli), Pseudomonas aeruginosa, and Acinetobacter baumannii were incubated with these flavin derivatives in vitro and were subsequently irradiated with visible light for seconds only. Singlet oxygen production in bacteria was proved by detecting its luminescence at 1270 nm. After irradiation, the number of viable bacteria decreased up to 6 log10 steps depending on the concentration of the flavin derivatives and the light dosimetry. The bactericidal effect of PIB was independent of the bacterial type and the corresponding antibiotic resistance pattern. In contrast, the photosensitizer concentration and light parameters used for bacteria killing did not affect cell viability of human keratinocytes (therapeutic window). Multiresistant bacteria can be safely and effectively killed by a combination of modified vitamin B2 molecules, oxygen and visible light, whereas normal skin cells survive. Further work will include these new photosensitizers for topical application to decolonize bacteria from skin and mucosa.

In this study, we investigated the utility of antimicrobial blue light therapy for multidrug-resistant Acinetobacter baumannii infection in a mouse burn model. A bioluminescent clinical isolate of multidrug-resistant A. baumannii was obtained. The susceptibility of A. baumannii to blue light (415 nm)-inactivation was compared in vitro to that of human keratinocytes. Repeated cycles of sublethal inactivation of bacterial by blue light were performed to investigate the potential resistance development of A. baumannii to blue light. A mouse model of third degree burn infected with A. baumannii was developed. A single exposure of blue light was initiated 30 minutes after bacterial inoculation to inactivate A. baumannii in mouse burns. It was found that the multidrug-resistant A. baumannii strain was significantly more susceptible than keratinocytes to blue light inactivation. Transmission electron microscopy revealed blue light-induced ultrastructural damage in A. baumannii cells. Fluorescence spectroscopy suggested that endogenous porphyrins exist in A. baumannii cells. Blue light at an exposure of 55.8 J/cm(2) significantly reduced the bacterial burden in mouse burns. No resistance development to blue light inactivation was observed in A. baumannii after 10 cycles of sublethal inactivation of bacteria. No significant DNA damage was detected in mouse skin by means of a skin TUNEL assay after a blue light exposure of 195 J/cm(2)., (© The Author 2014. Published by Oxford University Press on behalf of the Infectious Diseases Society of America. All rights reserved. For Permissions, please e-mail: journals.permissions@oup.com.)

One environmental concern related to hospital effluents is discharge of them without preliminary treatment. Antimicrobial photodynamic inactivation (PDI) may represent an alternative to the traditional expensive, unsafe and not always effective disinfection methods. The main goal of this work was to assess the efficiency of PDI on clinical multidrug-resistant (MDR) bacteria in hospital wastewaters in order to evaluate its potential use in treating hospital effluents. The efficiency of PDI was assessed using a cationic porphyrin as the photosensitizer (PS), four MDR bacteria either in phosphate buffered saline or in filtrated hospital wastewaters. The synergistic effect of PDI and antibiotics (ampicillin and chloramphenicol) was also evaluated, as well as the effect of the surfactant sodium dodecyl sulfate (SDS). The results show the efficient inactivation of MDR bacteria in PBS (reduction of 6-8 log after 270 min of irradiation at 40 W m(-2) with 5.0 μM of PS). In wastewater, the inactivation of the four MDR bacteria was again efficient and the decrease in bacterial survival starts even sooner. A faster decrease in bacterial survival occurred when PDI was combined with the addition of antibiotics, at sub-inhibitory and inhibitory concentrations, but the SDS did not affect the PDI efficiency. It can be concluded that PDI has potential to be an effective alternative for the inactivation of MDR bacteria in hospital wastewaters and that the presence of antibiotics may enhance its effectiveness.

Epidemics spread many types of pathogenic bacterial strains, especially strains of MRSA (Methicillin-resistant Staphylococcus aureus), which are being increasingly reported in many geographical areas [1]. This is becoming to be a serious global problem, particularly in hospitals. Not only are antibiotics proving to be increasingly ineffective but also the bacteria responsible for more than 70% of hospital-acquired bacterial infections are resistant to at least one of the drugs commonly used to treat them. In this study, hybrid coating A1 and nanocomposite hybrid coating A2 based on TMSPM (3-(trimethoxysilyl)propyl methacrylate, MMA (methyl methacrylate), TEOS (tetraethyl orthosilicate) and IPTI (titanium isopropoxide) containing silver and copper ions with or without nanoparticles of titanium dioxide were prepared by the sol-gel method. They were deposited on glass, poly(methyl methacrylate) and cotton using dip-coating or spin-coating, and then cured at 150 °C for 3 h or, in the case of poly(methyl methacrylate), at 100 °C for 4.5 h. The morphology and microstructure of these hybrid coatings were examined by SEM. The abrasion resistance was tested using a washability tester and found to depend heavily on the curing temperature. Seven types of bacterial strains were used to determine the profile of antibacterial activity, namely Escherichia coli, Staphylococcus aureus, Methicillin-resistant Staphylococcus aureus - MRSA (CCM 4223), MRSA-2 (CCM 7112), Acinetobacter baumanii, Pseudomonas aeruginosa, and Proteus vulgaris (according to ALE-G18, CSNI). All the samples were tested by irradiating with either a UV-A or a daylight fluorescent lamp. All types of hybrid coating A1 and nanocomposite hybrid coating A2 were found to possess an excellent antibacterial effect, including against the pathogenic bacterial strains of MRSA, which present a dangerous threat on a global scale., (Crown Copyright © 2012. Published by Elsevier B.V. All rights reserved.)

Background: Since the beginning of the conflicts in the Middle East, US Army physicians have noted a high rate of multidrug-resistant Acinetobacter baumannii infections among US soldiers wounded and initially treated in Iraq. In this study, we investigated the use of ultraviolet C (UVC) light for prevention of multidrug-resistant A. baumannii wound infections using mouse models., Methods: Partial-thickness skin abrasions and full-thickness burns in mice were infected with a multidrug-resistant A. baumannii isolate recovered from a wounded US soldier deployed to Iraq. The luxCDABE operon, which was contained in plasmid pMF 385, was cloned into the A. baumannii strain. This allowed real-time monitoring of the extent of infection in mice using bioluminescence imaging. UVC light was delivered to the mouse wounds at 30 minutes after the inoculation of A. baumannii. Groups of infected mouse wounds without being exposed to UVC served as the controls., Results: In vitro studies demonstrated that A. baumannii cells were inactivated at UVC exposures much lower than those needed for a similar effect on mammalian cells. It was observed in animal studies that UVC (3.24 J/cm(2) for abrasions and 2.59 J/cm(2) for burns) significantly reduced the bacterial burdens in UVC-treated wounds by approximately 10-fold compared with nontreated controls (p = 0.004 for abrasions, p = 0.019 for burns). DNA lesions were observed by immunofluorescence in mouse skin abrasions immediately after a UVC exposure of 3.24 J/cm(2); however, the lesions were extensively repaired within 72 hours., Conclusion: These results suggested that UVC may be useful in preventing combat-related wound infections.

Light is a ubiquitous environmental signal that many organisms sense and respond to by modulating their physiological responses accordingly. While this is an expected response among phototrophic microorganisms, the ability of chemotrophic prokaryotes to sense and react to light has become a puzzling and novel issue in bacterial physiology, particularly among bacterial pathogens. In this work, we show that the opportunistic pathogen Acinetobacter baumannii senses and responds to blue light. Motility and formation of biofilms and pellicles were observed only when bacterial cells were incubated in darkness. In contrast, the killing of Candida albicans filaments was enhanced when they were cocultured with bacteria under light. These bacterial responses depend on the expression of the A. baumannii ATCC 17978 A1S&#95;2225 gene, which codes for an 18.6-kDa protein that contains an N-terminal blue-light-sensing-using flavin (BLUF) domain and lacks a detectable output domain(s). Spectral analyses of the purified recombinant protein showed its ability to sense light by a red shift upon illumination. Therefore, the A1S&#95;2225 gene, which is present in several members of the Acinetobacter genus, was named blue-light-sensing A (blsA). Interestingly, temperature plays a role in the ability of A. baumannii to sense and respond to light via the BlsA photoreceptor protein.

Background and Objective: Phenothiazinium dyes have been reported to be effective photosensitizers inactivating a wide range of microorganisms in vitro after illumination with red light. However, their application in vivo has not extensively been explored. This study evaluates the bactericidal activity of phenothiazinium dyes against multidrug-resistant Acinetobacter baumannii both in vitro and in vivo., Study Design/materials and Methods: We report the investigation of toluidine blue O, methylene blue, 1,9-dimethylmethylene blue, and new methylene blue for photodynamic inactivation of multidrug-resistant A. baumannii in vitro. The most effective dye was selected to carry out in vivo studies using third-degree mouse burns infected with a bioluminescent A. baumannii strain, upon irradiation with a 652 nm noncoherent light source. The mice were imaged daily for 2 weeks to observe differences in the bioluminescence-time curve between the photodynamic therapy (PDT)-treated mice in comparison with untreated burns., Results: All the dyes were effective in vitro against A. baumannii after 30 J/cm(2) irradiation of 635 or 652 nm red light had been delivered, with more effective killing when the dye remained in solution. New methylene blue was the most effective of the four dyes, achieving a 3.2-log reduction of the bacterial luminescence during PDT in vivo after 360 J/cm(2) and an 800 microM dye dose. Moreover, a statistically significant reduction of the area under the bioluminescence-time curve of PDT-treated mice was observed showing that the infection did not recur after PDT., Conclusions: Phenothiazinium dyes, and especially new methylene blue, are potential photosensitizers for PDT to treat burns infected with multidrug-resistant A. baumannii in vivo.

Multidrug-resistant Acinetobacter baumannii infections represent a growing problem, especially in traumatic wounds and burns suffered by military personnel injured in Middle Eastern conflicts. Effective treatment with traditional antibiotics can be extremely difficult, and new antimicrobial approaches are being investigated. One of these alternatives to antimicrobials could be the combination of nontoxic photosensitizers (PSs) and visible light, known as photodynamic therapy (PDT). We report on the establishment of a new mouse model of full-thickness thermal burns infected with a bioluminescent derivative of a clinical Iraqi isolate of A. baumannii and its PDT treatment by topical application of a PS produced by the covalent conjugation of chlorin(e6) to polyethylenimine, followed by illumination of the burn surface with red light. Application of 10(8) A. baumannii cells to the surface of 10-s burns made on the dorsal surface of shaved female BALB/c mice led to chronic infections that lasted, on average, 22 days and that were characterized by a remarkably stable bacterial bioluminescence. PDT carried out on day 0 soon after application of the bacteria gave over 3 log units of loss of bacterial luminescence in a light exposure-dependent manner, while PDT carried out on day 1 and day 2 gave an approximately 1.7-log reduction. The application of PS dissolved in 10% or 20% dimethyl sulfoxide without light gave only a modest reduction in the bacterial luminescence from mouse burns. Some bacterial regrowth in the treated burn was observed but was generally modest. It was also found that PDT did not lead to the inhibition of wound healing. The data suggest that PDT may be an effective new treatment for multidrug-resistant localized A. baumannii infections.

The antibacterial activity of photocatalytic titanium dioxide (TiO(2)) substrates is induced primarily by UV light irradiation. Recently, nitrogen- and carbon-doped TiO(2) substrates were shown to exhibit photocatalytic activities under visible-light illumination. Their antibacterial activity, however, remains to be quantified. In this study, we demonstrated that nitrogen-doped TiO(2) substrates have superior visible-light-induced bactericidal activity against Escherichia coli compared to pure TiO(2) and carbon-doped TiO(2) substrates. We also found that protein- and light-absorbing contaminants partially reduce the bactericidal activity of nitrogen-doped TiO(2) substrates due to their light-shielding effects. In the pathogen-killing experiment, a significantly higher proportion of all tested pathogens, including Shigella flexneri, Listeria monocytogenes, Vibrio parahaemolyticus, Staphylococcus aureus, Streptococcus pyogenes, and Acinetobacter baumannii, were killed by visible-light-illuminated nitrogen-doped TiO(2) substrates than by pure TiO(2) substrates. These findings suggest that nitrogen-doped TiO(2) has potential application in the development of alternative disinfectants for environmental and medical usages.