Photodynamic Inactivation of Foodborne Bacteria: Screening of 32 Potential Photosensitizers.

The development of novel antimicrobial technologies for the food industry represents an important strategy to improve food safety. Antimicrobial photodynamic disinfection (aPDD) is a method that can inactivate microbes without the use of harsh chemicals. aPDD involves the administration of a non-toxic, light-sensitive substance, known as a photosensitizer, followed by exposure to visible light at a specific wavelength. The objective of this study was to screen the antimicrobial photodynamic efficacy of 32 food-safe pigments tested as candidate photosensitizers (PSs) against pathogenic and food-spoilage bacterial suspensions as well as biofilms grown on relevant food contact surfaces. This screening evaluated the minimum bactericidal concentration (MBC), minimum biofilm eradication concentration (MBEC), and colony forming unit (CFU) reduction against Salmonella enterica, methicillin-resistant Staphylococcus aureus (MRSA), Pseudomonas fragi, and Brochothrix thermosphacta. Based on multiple characteristics, including solubility and the ability to reduce the biofilms by at least 3 log10 CFU/sample, 4 out of the 32 PSs were selected for further optimization against S. enterica and MRSA, including sunset yellow, curcumin, riboflavin-5'-phosphate (R-5-P), and erythrosin B. Optimized factors included the PS concentration, irradiance, and time of light exposure. Finally, 0.1% w/v R-5-P, irradiated with a 445 nm LED at 55.5 J/cm2, yielded a "max kill" (upwards of 3 to 7 log10 CFU/sample) against S. enterica and MRSA biofilms grown on metallic food contact surfaces, proving its potential for industrial applications. Overall, the aPDD method shows substantial promise as an alternative to existing disinfection technologies used in the food processing industry.

New Insights into the Bacterial Targets of Antimicrobial Blue Light.

Antimicrobial blue light (aBL) offers efficacy and safety in treating infections. However, the bacterial targets for aBL are still poorly understood and may be dependent on bacterial species. Here, we investigated the biological targets of bacterial killing by aBL (λ = 410 nm) on three pathogens: Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. Initially, we evaluated the killing kinetics of bacteria exposed to aBL and used this information to calculate the lethal doses (LD) responsible for killing 90 and 99.9% of bacteria. We also quantified endogenous porphyrins and assessed their spatial distribution. We then quantified and suppressed reactive oxygen species (ROS) production in bacteria to investigate their role in bacterial killing by aBL. We also assessed aBL-induced DNA damage, protein carbonylation, lipid peroxidation, and membrane permeability in bacteria. Our data showed that P. aeruginosa was more susceptible to aBL (LD99.9 = 54.7 J/cm2) relative to S. aureus (LD99.9 = 158.9 J/cm2) and E. coli (LD99.9 = 195 J/cm2). P. aeruginosa exhibited the highest concentration of endogenous porphyrins and level of ROS production relative to the other species. However, unlike other species, DNA degradation was not observed in P. aeruginosa. Sublethal doses of blue light (LD99.9). We conclude that the primary targets of aBL depend on the species, which are probably driven by variable antioxidant and DNA-repair mechanisms. IMPORTANCE Antimicrobial-drug development is facing increased scrutiny following the worldwide antibiotic crisis. Scientists across the world have recognized the urgent need for new antimicrobial therapies. In this sense, antimicrobial blue light (aBL) is a promising option due to its antimicrobial properties. Although aBL can damage different cell structures, the targets responsible for bacterial inactivation have still not been completely established and require further exploration. In our study, we conducted a thorough investigation to identify the possible aBL targets and gain insights into the bactericidal effects of aBL on three relevant pathogens: Staphylococcus aureus, Escherichia coli, and Pseudomonas aeruginosa. This research not only adds new content to blue light studies but opens new perspectives to antimicrobial applications.

The Biochemical Mechanisms of Antimicrobial Photodynamic Therapy†.

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.

A systematic scoping review of ultraviolet C (UVC) light systems for SARS-CoV-2 inactivation.

A significant amount of epidemiological evidence has underlined that human-to-human transmission due to close contacts is considered the main pathway of transmission, however since the SARS-CoV-2 can also survive in aerosols, water, and surfaces, the development and implementation of effective decontamination strategies are urgently required. In this regard, ultraviolet germicidal irradiation (UVGI) using ultraviolet C (UVC) has been proposed to disinfect different environments and surfaces contaminated by SARS-CoV-2. Herein, we performed a systematic scoping review strictly focused on peer-reviewed studies published in English that reported experimental results of UVC-based technologies against the SARS-CoV-2 virus. Studies were retrieved from PubMed and the Web of Science database. After our criterious screening, we identified 13 eligible articles that used UVC-based systems to inactivate SARS-CoV-2. We noticed the use of different UVC wavelengths, technologies, and light doses. The initial viral titer was also heterogeneous among studies. Most studies reported virus inactivation in well plates, even though virus persistence on N95 respirators and different surfaces were also evaluated. SARS-CoV-2 inactivation reached from 90% to 100% depending on experimental conditions. We concluded that there is sufficient evidence to support the use of UVC-based technologies against SARS-CoV-2. However, appropriate implementation is required to guarantee the efficacy and safety of UVC strategies to control the COVID-19 pandemic.

Methylene blue-mediated antimicrobial photodynamic therapy can be a novel non-antibiotic platform for bovine digital dermatitis.

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/cm2, 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.

Antimicrobial blue light and photodynamic therapy inhibit clinically relevant ß-lactamases with extended-spectrum (ESBL) and carbapenemase activity.

INTRODUCTION: The production of ß-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 ß-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 ß-lactamase enzymes assigned to the A, B and D Ambler molecular classes. Furthermore, ß-lactamase activity inhibition mediated by antimicrobial blue light and methylene blue-mediated antimicrobial photodynamic therapy was measured by using the chromogenic ß-lactam substrate nitrocefin. RESULTS: ß-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 ß-lactamases, including extended-spectrum ß-lactamases and carbapenemases.

Light-based technologies for management of COVID-19 pandemic crisis.

The global dissemination of the novel coronavirus disease (COVID-19) has accelerated the need for the implementation of effective antimicrobial strategies to target the causative agent SARS-CoV-2. Light-based technologies have a demonstrable broad range of activity over standard chemotherapeutic antimicrobials and conventional disinfectants, negligible emergence of resistance, and the capability to modulate the host immune response. This perspective article identifies the benefits, challenges, and pitfalls of repurposing light-based strategies to combat the emergence of COVID-19 pandemic.

Hypervirulent and hypermucoviscous strains of Klebsiella pneumoniae challenged by antimicrobial strategies using visible light.

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.

Preclinical Investigation of Methylene Blue-mediated Antimicrobial Photodynamic Therapy on Leishmania Parasites Using Real-Time Bioluminescence.

Cutaneous leishmaniasis (CL) is a neglected disease that promotes destructive lesions. Difficulties in treatment are related to accessibility of drugs, resistance and toxicity. Antimicrobial photodynamic therapy (APDT) has been emerging as a promising treatment for CL. In this work, we evaluated methylene blue (MB)-mediated APDT (MB-APDT) on Leishmania amazonensis in vitro and in vivo by bioluminescence technique. In vitro, MB-APDT was performed using a red LED (λ = 660 ± 11 nm, 100 mW cm-2 ) and MB (100 µm) at different light doses. In vivo, mice were infected and 4 weeks later, randomly divided into three groups: control, APDT 1 (single session) and APDT 2 (two sessions of MB-APDT). MB was used at 100 µm and energy dose was established at 150 J cm-2 . Parasite burden, lesion size and pain were evaluated weekly for 4 weeks. In vitro, lethal dose for 90% parasite inactivation was achieved at 48.8 J cm-2 . In vivo, although APDT 1 and APDT 2 groups have showed similar parasite burden after 4 weeks, two sessions were clinically better, especially considering the inflammatory process associated to CL. Our findings reinforce MB-APDT as a cost-effective treatment to combat CL.

Identification and genomic features of halotolerant extended-spectrum-ß-lactamase (CTX-M)-producing Escherichia coli in urban-impacted coastal waters, Southeast Brazil.

We report the occurrence and genomic analysis of extended-spectrum ß-lactamase (CTX-M)-producing Escherichia coli in anthropogenically polluted coastal waters of Southeast Brazil. E. coli strains belonging to sequence types (STs) ST10, ST38, ST155 and ST1284 exhibited a wide resistome, with genes conferring resistance to medically relevant antimicrobials and heavy metals, and a halophilic behavior (tolerance to 9-10% NaCl). These findings suggest a heavy contamination in this area by critical priority bacteria adapted to marine environments, which might have negative impacts on human and ocean health.

Inactivation kinetics and lethal dose analysis of antimicrobial blue light and photodynamic therapy.

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.

Antimicrobial blue light inactivation of international clones of multidrug-resistant Escherichia coli ST10, ST131 and ST648.

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, blaCTX-M or blaKPC-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 ß-lactam antibiotics.

Effective treatment and decolonization of a dog infected with carbapenemase (VIM-2)-producing Pseudomonas aeruginosa using probiotic and photodynamic therapies.

BACKGROUND: Carbapenem-resistant bacterial infections are a critical problem in veterinary medicine with limited treatment options. OBJECTIVE: To describe effective probiotic and photodynamic therapy of a dog with gut colonization and ear infection caused by a hospital-associated lineage of carbapenemase (VIM-2)-producing Pseudomonas aeruginosa. ANIMALS: A 5-year-old Lhasa apso dog presented with otitis externa. METHODS AND MATERIALS: Unilateral otitis externa caused by carbapenem-resistant P. aeruginosa was treated with antimicrobial photodynamic therapy (aPDT) using methylene blue as photosensitizer [wavelength 660 nm, fluence 140 J/cm2 , 8 J and 80 s per point (six equidistant points), 100 mW, spot size 0.028 cm2 and fluence rate 3.5 W/cm2 ]. The isolated bacterial strain also was tested for susceptibility to in vitro aPDT where the survival fraction was quantified by colony forming unit counts after exposure to increasing light doses. For decolonization, probiotic supplements were orally administered (once daily) for 14 days. Effectiveness of probiotics and photodynamic therapy was evaluated by clinical and microbiological culture assays. RESULTS: Complete resolution of clinical signs was achieved by Day 7 after aPDT. Samples collected immediately and after seven and 14 days following aPDT were negative for VIM-2-producing P. aeruginosa. Oral and rectal swabs collected on days 7, 14 and 21 after probiotic therapy, confirmed effective gastrointestinal decolonization. CONCLUSIONS AND CLINICAL IMPORTANCE: Combined use of aPDT and probiotics could be a promising therapeutic strategy for treatment of superficial infections produced by carbapenem-resistant bacteria, while avoiding recurrent infection due to intestinal bacterial carriage of these multidrug-resistant pathogens.

Photodynamic damage predominates on different targets depending on cell growth phase of Candida albicans.

Photodynamic inactivation (PDI) has been reported to be effective to eradicate a wide variety of pathogens, including antimicrobial-resistant microorganisms. The aim of this study was to identify the potential molecular targets of PDI depending on growth phase of Candida albicans. Fungal cells in lag (6h) and stationary (48h) phases were submitted to PDI mediated by methylene blue (MB) combined with a (662±21) nm-LED, at 360mW of optical power. Pre-irradiation time was 10min and exposure times were 12min, 15min and 18min delivering radiant exposures of 129.6J/cm2, 162J/cm2 and 194.4J/cm2, respectively, on a 24-well plate of about 2cm2 at an irradiance of 180mW/cm2. Scanning electron microscopy (SEM), transmission electron microscopy (TEM), atomic force spectroscopy (AFS) and Fourier transform infrared spectroscopy (FT-IR) were employed to evaluate the photodynamic effect in young and old fungal cells following 15min of irradiation. Morphological analysis revealed wrinkled and shrunk fungal cell membrane for both growth phases while extracellular polymeric substance (EPS) removal was only observed for old fungal cells. Damaged intracellular structures were more pronounced in young fungal cells. The surface nanostiffness of young fungal cells decreased after PDI but increased for old fungal cells. Cellular adhesion force was reduced for both growth phases. Fungal cells in lag phase predominantly showed degradation of nucleic acids and proteins, while fungal cells in stationary phase showed more pronounced degradation of polysaccharides and lipids. Taken together, our results indicate different molecular targets for fungal cells in lag and stationary growth phase following PDI.

Detection of Colistin-Resistant MCR-1-Positive Escherichia coli by Use of Assays Based on Inhibition by EDTA and Zeta Potential.

The emergence and rapid dissemination of colistin-resistant Escherichia coli carrying the plasmid-mediated mcr-1 gene have created an urgent need to develop specific screening methods. In this study, we evaluated four assays based on the inhibition of MCR-1 activity by EDTA: (i) a combined-disk test (CDT) comparing the inhibition zones of colistin and colistin (10 µg) plus EDTA (100 mM); (ii) reduction of colistin MIC (CMR) in the presence of EDTA (80 µg/ml); (iii) a modified rapid polymyxin Nordmann/Poirel test (MPNP); and (iv) alteration of zeta potential (RZP = ZP+EDTA/ZP-EDTA). We obtained encouraging results for the detection of MCR-1 in E. coli isolates recovered from human, food, and animal samples, using the following assay parameters: ≥3 mm difference in the inhibition zones between colistin disks without and with EDTA; ≥4-fold colistin MIC decrease in the presence of EDTA; RZP of ≥2.5; and the absence of metabolic activity and proliferation, indicated by unchanged color of phenol red in the presence of colistin-EDTA, in the MPNP test. In this regard, the CDT, CMR, RZP, and MPNP assays exhibited sensitivities of 96.7, 96.7, 95.1, and 96.7% and specificities of 89.6, 83.3, 100, and 100%, respectively, for detecting MCR-1-positive E. coli Our results demonstrate that inhibition by EDTA and zeta potential assays may provide simple and inexpensive methods for the presumptive detection of MCR-1-producing E. coli isolates in human and veterinary diagnostic laboratories.

Colistin-Resistant mcr-1-Positive Escherichia coli on Public Beaches, an Infectious Threat Emerging in Recreational Waters.

The emergence and rapid spread of colistin-resistant Escherichia coli carrying the mcr-1 gene have generated an urgent need to strengthen surveillance. We performed a meticulous investigation of strains of this sort, which resulted in the identification of international clones of E. coli carrying IncX4-plasmid-mediated mcr-1 and blaCTX-M genes in recreational waters of public urban beaches in cities with high tourist turnover, highlighting a new environmental reservoir.

The optical properties of mouse skin in the visible and near infrared spectral regions.

Visible and near-infrared radiation is now widely employed in health science and technology. Pre-clinical trials are still essential to allow appropriate translation of optical methods into clinical practice. Our results stress the importance of considering the mouse strain and gender when planning pre-clinical experiments that depend on light-skin interactions. Here, we evaluated the optical properties of depilated albino and pigmented mouse skin using reproducible methods to determine parameters that have wide applicability in biomedical optics. Light penetration depth (δ), absorption (µa), reduced scattering (µ's) and reduced attenuation (µ't) coefficients were calculated using the Kubelka-Munk model of photon transport and spectrophotometric measurements. Within a broad wavelength coverage (400-1400nm), the main optical tissue interactions of visible and near infrared radiation could be inferred. Histological analysis was performed to correlate the findings with tissue composition and structure. Disperse melanin granules present in depilated pigmented mouse skin were shown to be irrelevant for light absorption. Gender mostly affected optical properties in the visible range due to variations in blood and abundance of dense connective tissue. On the other hand, mouse strains could produce more variations in the hydration level of skin, leading to changes in absorption in the infrared spectral region. A spectral region of minimal light attenuation, commonly referred as the "optical window", was observed between 600 and 1350nm.

CdTe quantum dots conjugated to concanavalin A as potential fluorescent molecular probes for saccharides detection in Candida albicans.

Semiconductor colloidal quantum dots (QDs) have been applied in biological analysis due to their unique optical properties and their versatility to be conjugated to biomolecules, such as lectins and antibodies, acquiring specificity to label a variety of targets. Concanavalin A (Con A) lectin binds specifically to α-d-mannose and α-d-glucose regions of saccharides that are usually expressed on membranes of mammalian cells and on cell walls of microbials. Candida albicans is the most common fungal opportunistic pathogen present in humans. Therefore, in this work, this fungus was chosen as a model for understanding cells and biofilm-forming organisms. Here, we report an efficient bioconjugation process to bind CdTe (Cadmium Telluride) QDs to Con A, and applied the bioconjugates to label saccharide structures on the cellular surface of C. albicans suspensions and biofilms. By accomplishing hemagglutination experiments and circular dichroism, we observed that the Con A structure and biochemical properties were preserved after the bioconjugation. Fluorescence microscopy images of yeasts and hyphae cells, as well as biofilms, incubated with QDs-(Con A) showed a bright orange fluorescence profile, indicating that the cell walls were specifically labeled. Furthermore, flow cytometry measurements confirmed that over 93% of the yeast cells were successfully labeled by QD-(Con A) complex. In contrast, non-conjugated QDs or QDs-(inhibited Con A) do not label any kind of biological system tested, indicating that the bioconjugation was specific and efficient. The staining pattern of the cells and biofilms demonstrate that QDs were effectively bioconjugated to Con A with specific labeling of saccharide-rich structures on C. albicans. Consequently, this work opens new possibilities to monitor glucose and mannose molecules through fluorescence techniques, which can help to optimize phototherapy protocols for this kind of fungus.