Search

The unbridled dissemination of multidrug-resistant pathogens is a major threat to global health and urgently demands novel therapeutic alternatives. Antimicrobial photodynamic therapy (aPDT) has been developed as a promising approach to treat localized infections regardless of drug resistance profile or taxonomy. Even though this technique has been known for more than a century, discussions and speculations regarding the biochemical mechanisms of microbial inactivation have never reached a consensus on what is the primary cause of cell death. Since photochemically generated oxidants promote ubiquitous reactions with various biomolecules, researchers simply assumed that all cellular structures are equally damaged. In this study, biochemical, molecular, biological and advanced microscopy techniques were employed to investigate whether protein, membrane or DNA damage correlates better with dose-dependent microbial inactivation kinetics. We showed that although mild membrane permeabilization and late DNA damage occur, no correlation with inactivation kinetics was found. On the other hand, protein degradation was analyzed by three different methods and showed a dose-dependent trend that matches microbial inactivation kinetics. Our results provide a deeper mechanistic understanding of aPDT that can guide the scientific community toward the development of optimized photosensitizing drugs and also rationally propose synergistic combinations with antimicrobial chemotherapy., (© 2022 American Society for Photobiology.)

Dermatophytosis is a superficial skin infection that widely effects companion animals. Miscrosporum canis is one of the most prevalent species isolated from dogs and cats, and because of the serious zoonotic potential, short-term treatment regimens are preferred to prevent the spread of disease either by direct contact or through contamination of the environment. Antimicrobial photodynamic therapy (APDT) has emerged as a promising strategy able to kill effectively a wide range of pathogens in a short period with minimal morbidity . In this case report, a 7-year-old male dog was diagnosed with dermatophytosis caused by M. canis. Methylene blue-mediated antimicrobial photodynamic therapy (MB-APDT) was applied over the lesions in two sessions with an interval of 7 days. The dog successfully healed, achieving a complete clinical cure after 21 days, without reports of recurrence after a follow-up period of 6 months. Therefore, MB-APDT could be a potential ally of small animal clinicians to treat superficial fungal diseases and should be further explored in Veterinary Medicine., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Bovine digital dermatitis (BDD) is one of the most important diseases that effect dairy cows. Methylene blue-mediated antimicrobial photodynamic therapy (MB-APDT) emerges as a promising technique to treat superficial infections in bovines., Methods: Twenty BDD lesions located at the skin horn transition of the claw of pelvic limbs of 16 cows were treated by MB-APDT, using a red LED cluster (λ = 660 nm, irradiance =60 mW/cm 2 , exposure time = 40 s) combined with topical application of MB at 0.01 %; or by topical application of OXY (500 mg in 20 % solution). Each lesion was treated twice with an interval of 14 days. Lesions were weekly evaluated until day 28 by clinical analysis and by histological examination on days 0 and 28., Results: Both treatments led to a similar reduction of lesions area. At day 28, three lesions treated by OXY did not present completely recovery, whereas no lesions were observed in MB-APDT group. OXY resulted in a slight increase in type I and III collagen levels, while MB-APDT led to a significant increase in the total area of both collagen types. An abundant number of spirochetes were histologically observed in all lesions before treatments. On the 28th day, five lesions treated by OXY still presented a slight number of spirochetes, whereas in MB-APDT group no spirochetes were evidenced., Conclusion: Our findings suggest that MB-APDT is more effective than OXY and could be used in Veterinary practice to fight BDD., (Copyright © 2021 Elsevier B.V. All rights reserved.)

Background: Rumenostomy is a useful procedure commonly performed in cattle for medical treatment of domestic ruminants with forestomach diseases. Methylene blue (MB)-mediated antimicrobial photodynamic therapy (APDT) has been broadly investigated to treat infected wounds., Aim: The aim of the study was to evaluate the effectiveness of MB-mediated APDT (MB-APDT) combined with chlorhexidine and zinc oxide ointment on wound healing process after rumenostomy., Methods: Fourteen Nelore bulls were subjected to rumenostomy procedure. Animals were randomly divided into MB-APDT (MB associated with a red diode laser performed immediately after surgery and repeated on days 3, 5, 7 and 10) and control groups. Daily care included topical cleaning with chlorhexidine 2% followed by topical zinc oxide ointment. Animals were followed-up until the 28th day., Results: Wounds presented a better post-surgical profile in MB-APDT group when it was compared with the control group. In MB-APDT group, it was also possible to observe less pain on palpation of wounds borders, less edema and inflammatory exudate. Additionally, animals from MB-APDT group were faster discharged from the cattle care facility., Conclusion: Our results support the use of MB-APDT for the post-surgical management of rumenostomy. This pilot study ratifies the use of APDT in cattle and also suggests that it could be performed for other surgical procedures as a complementary approach or an alternative for topical administration of antibiotics., (Copyright © 2020. Published by Elsevier B.V.)

Introduction: The production of β-lactamases by Gram-negative bacteria is among the most important factors of resistance to antibiotics, which has contributed to therapeutic failures that currently threaten human and veterinary medicine worldwide. Antimicrobial photodynamic therapy and antimicrobial blue light have a broad-spectrum antibacterial activity against multidrug-resistant and hypervirulent pathogens., Objective: To investigate the ability of antimicrobial blue light to inhibit the hydrolytic activity of clinically relevant β-lactamase enzymes (i.e., KPC, IMP, OXA, CTX-M, and SHV), with further comparison of the inhibitory effects of antimicrobial blue light with methylene blue-mediated antimicrobial photodynamic therapy., Methods: Blue LED light (λ = 410 ± 10 nm) alone or red LED light (λ = 660 ± 10 nm) in combination with methylene blue were used to inactivate, in vitro, suspensions of Klebsiella pneumoniae strains producing clinically important β-lactamase enzymes assigned to the A, B and D Ambler molecular classes. Furthermore, β-lactamase activity inhibition mediated by antimicrobial blue light and methylene blue-mediated antimicrobial photodynamic therapy was measured by using the chromogenic β-lactam substrate nitrocefin., Results: β-lactamase activities were effectively inactivated by both visible light-based approaches. In this regard, antimicrobial blue light and methylene blue-antimicrobial photodynamic therapy led to a significant reduction in the hydrolysis of nitrocefin (81-98 %)., Conclusion: Sublethal doses of antimicrobial blue light and methylene blue-mediated antimicrobial photodynamic therapy are equally effective to inhibit clinically significant β-lactamases, including extended-spectrum β-lactamases and carbapenemases., (Copyright © 2020 Elsevier B.V. All rights reserved.)

Introduction: Infections caused by hypervirulent and/or hypermucoviscous Klebsiella pneumoniae (K. pneumoniae) strains are frequently reported worldwide. Since convergence of hypervirulence and drug-resistance emerged as a serious clinical problem, novel therapeutic strategies are worthy of investigation. In this regard, antimicrobial photodynamic therapy and blue light have proven to be effective against a broad-spectrum of clinically relevant pathogens but have never been tested for hypervirulent/hypermucoviscous strains. Thus, this study investigated the influence of hypermucoviscosity and hypervirulence over the photoinactivation efficacy of blue light alone or antimicrobial photodynamic therapy mediated by methylene blue and red light., Methods: Five clinical isolates of K. pneumoniae were screened for hypermucoviscosity by string test and for hypervirulence by a Galleria mellonella model of systemic infection. Strains were then challenged by both photoinactivation methods performed in vitro. All tests also included a non-hypervirulent/hypermucoviscous control strain for comparison., Results: All K. pneumoniae strains were effectively inactivated by both light-based antimicrobial strategies. Hypervirulent/hypermucoviscous strains exposed to photodynamic therapy presented rapid and consistent inactivation kinetics, while blue light led to slower and more variable inactivation kinetics., Conclusion: Hypermucoviscosity and hypervirulence does not confer tolerance in K. pneumoniae against photoinactivation. Antimicrobial photodynamic therapy represents an interesting alternative to treat localised infections because it is a fast procedure with high effectiveness. On the other hand, antimicrobial blue light could be used to decontaminate hospital environments since no photosensitiser administration is required and harmful effects of ultraviolet light are avoided. Therefore, visible light-based strategies present great potential for the development of safe and effective antimicrobial technologies against such aggressive pathogens., (Copyright © 2020 Elsevier Ltd. All rights reserved.)

Background: Antimicrobial Photodynamic therapy (A-PDT) has been used to treat infections. Currently, microbial inactivation data is reported presenting survival fraction averages and standard errors as discrete points instead of a continuous curve of inactivation kinetics. Standardization of this approach would allow clinical protocols to be introduced globally, instead of the piecemeal situation which currently applies., Methods: To this end, we used a power-law function to fit inactivation kinetics and directly report values of lethal doses (LD) and a tolerance factor (T) that informs if inactivation rate varies along the irradiation procedure. A deduced formula was also tested to predict LD for any given survival fraction value. We analyzed the photoantimicrobial effect caused by red light activation of methylene blue (MB-APDT) and by blue light (BL) activation of endogenous microbial pigments against 5 clinically relevant pathogens., Results: Following MB- APDT, Escherichia coli and Staphylococcus aureus cells become increasingly more tolerant to inactivation along the irradiation process (T < 1). Klebsiella pneumoniae presents opposite behavior, i.e., more inactivation is observed towards the end of the process (T > 1). P. aeruginosa and Candida albicans present constant inactivation rate (T˜1). In contrast, all bacterial species presented similar behavior during inactivation caused by BL, i.e., continuously becoming more sensitive to blue light exposure (T > 1)., Conclusion: The power-law function successfully fit all experimental data. Our proposed method precisely predicted LD and T values. We expect that these analytical models may contribute to more standardized methods for comparisons of photodynamic inactivation efficiencies., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Background: International clones of multidrug-resistant Escherichia coli have been a leading cause of human and animal infections worldwide. Microbial inactivation by blue light has been proposed as an effective treatment for superficial infections and surface contamination., Aim: To evaluate the inactivation efficacy of blue light irradiation against high-risk multidrug-resistant strains of E. coli., Methods: Blue LED light (λ = 410 nm) was used to inactivate in vitro suspensions of colistin- broad-spectrum cephalosporin-, or carbapenem-resistant E. coli strains belonging to sequence types (STs) ST10, ST131 and ST648, carrying mcr-1, bla CTX-M or bla KPC-2 genes., Results: Our results showed that all E. coli strains were susceptible to blue light irradiation, independently of antibiotic resistance and virulence profiles. In addition, blue light irradiation induced a strain-specific and dose-dependent bacterial effect., Conclusion: Our results support use of blue light as a promising antimicrobial option against MDR pathogens, including high-risk clones of E. coli displaying resistance to polymyxins or broad-spectrum β-lactam antibiotics., (Copyright © 2019 Elsevier B.V. All rights reserved.)

Prototheca spp. are pathogenic algae with important zoonotic potential. Most importantly, these algae often infect dairy cattle. Since there is no effective therapy against the algae, the standard recommendation is the disposal or culling of infected cows to avoid outbreaks. This study investigated the ability of blue light to inactivate pathogenic Prototheca species. Blue LED light (λ = 410 nm) was used to inactivate in vitro suspensions of P. zopfii genotypes 1 and 2, and P. blaschkeae. Our results showed that blue light irradiation induced a strain-specific dose-dependent algicidal effect against all tested strains. P. zopfii genotype 1, was more sensitive than genotype 2 and P. blaschkeae was the most tolerant. Even though we observed different inactivation kinetics, all strains presented significant photoinactivation levels within feasible procedure periods. Therefore, we conclude that blue light irradiation offers promising potential for the development of novel technologies that control contaminations and infections caused by Prototheca spp., (Copyright © 2019. Published by Elsevier B.V.)

Background: Antimicrobial photodynamic therapy (APDT) has been broadly investigated as an alternative to treat localized infections, without leading to the selection of resistant microorganisms. Infectious stomatitis is a multifactorial disease frequently reported in captive snakes characterized by infection of the oral mucosa and surrounding tissues. In this study, we investigated methylene blue (MB)-mediated APDT to treat infectious stomatitis in snakes and verified the resistance phenotype and genotype before and after APDT., Methods: Three Boid snakes presented petechiae, edema and caseous material in their oral cavities. MB (0.01%) was applied on the lesions and after 5min they were irradiated using a red laser (λ=660nm), fluence of 280J/cm 2 , 8J and 80s per point, 100mW, spot size 0.028cm 2 and fluence rate of 3.5W/cm 2 . APDT was repeated once a week during 3 months. Samples of the lesions were collected to identify bacteria and antibiotic resistance profiles. To analyze the clonality of bacterial isolates before and after APDT, isolates were subjected to ERIC PCR analysis., Results: Snakes presented clinical improvement such as reduction of inflammatory signs and caseous material. Pseudomonas aeruginosa and Escherichia coli were present in all snakes; Klebsiella pneumoniae and Morganella morganii were also identified in some animals. We also observed that the oral microbiota was completely replaced following APDT. However, K. pneumoniae isolates before and after APDT were a single clone with 100% of genetic similarity that lost resistance phenotype for seven antibiotics of four classes., Conclusions: These results show that APDT can be used to treat infectious stomatitis in snakes., (Copyright © 2017 Elsevier B.V. All rights reserved.)

Background: Bovine mastitis is considered the most important disease of worldwide dairy industry. Treatment of this disease is based on the application intramammary antibiotic, which favors an increase in the number of resistant bacteria in the last decade. Photodynamic inactivation (PDI) has been investigated in different areas of Health Sciences, and has shown great potential for inactivating different pathogens, without any selection of resistant microorganisms. The objective of this study was to investigate the efficacy of PDI in the inactivation of pathogens associated with bovine mastitis., Methods: We tested the effectiveness of PDI against antibiotic resistant strains, isolated from bovine mastitis, from the following species: Staphylococcus aureus, Streptococcus agalactiae, Streptococcus dysgalactiae, Corynebacterium bovis, and the alga Prototheca zopfii. Nine experimental groups were evaluated: control, no treatment; light only, irradiation of a red light-emitting diode (λ=662 (20) nm) for 180 s; exposure to 50 μM methylene blue alone for 5 min; and PDI for 5, 10, 30, 60, 120 and 180 s., Results: S. dysgalactiae, S. aureus, and C. bovis were inactivated after 30s of irradiation, whereas S. agalactiae was inactivated after 120 s and P. zopfii at 180 s of irradiation., Conclusion: These results show that PDI can be an interesting tool for inactivating pathogens for bovine mastitis., (Copyright © 2015 Elsevier B.V. All rights reserved.)

Background: Bumblefoot, referring to bed-sore-like foot lesions, is one of the most important clinical complications in captive birds and has a multifactorial etiology. Photodynamic therapy has been proposed as an alternative treatment for localized infections in response to the escalating problem of antibiotic resistance. The aim of this study was to compare outcomes in a group of captive Spheniscus magellanicus with bumblefoot lesions treated with photodynamic therapy (PDT) or antibiotics (ATB)., Methods: Ten captive Magellanic penguins with preexisting stage III bumblefoot lesions were selected and randomly divided into one PDT and one ATB group, each including 11 pelvic-limb lesions. All animals underwent surgical debridement of lesions. In the ATB group, antibiotic ointment was applied topically three times a week, and systemic antibiotic and anti-inflammatory drugs were administered daily. In the PDT group, photodynamic therapy was applied three times a week without the use of topical or systemic medication. Lesion areas were photographed, and swabs were collected for culture and sensitivity, on the first day and every 14 days for a total of 84 days. The four species of bacteria showing the most resistance to the antibiotics screened on the antibiogram were used to determine resistance to PDT with an in vitro test., Results: There were significant differences in healing rate and average healing time between the PDT and ATB groups (63.62% vs. 9.09% and 42 vs. 70 days, respectively)., Conclusion: The findings of this study attest to the effectiveness of photodynamic therapy for the treatment of stage III bumblefoot in Spheniscus magellanicus., (Copyright © 2015 The Authors. Published by Elsevier B.V. All rights reserved.)

Pododermatitis is currently one of most frequent and important clinical complications in seabirds kept in captivity or in rehabilitation centers. In this study, five Magellanic penguins with previous pododermatitis lesions on their footpad were treated with photodynamic therapy (PDT). All PDT treated lesions successfully regressed and no recurrence was observed during the 6-month follow-up period. PDT seems to be an inexpensive and effective alternative treatment for pododermatitis in Magellanic penguins encouraging further research on this topic., (© 2014 Wiley Periodicals, Inc.)