Name: Forfatter Spotlight: Fotodynamisk terapi som en ny tilnærming for å indusere små kolonier i medikamentresistent Candida for soppdrepende forskning
Uploaded: 2024-03-29T14:30:00
Description: Watch this Scientific Journal Video about Photodynamic Therapy as a Novel Approach to Induce Petite Colonies in Drug-Resistant Candida for Antifungal Research at JoVE.com

Dette arbeidet har mottatt finansiering fra departementet for vitenskap og teknologi, Taiwan [MOST 110-2314-B-006-086-MY3], National Cheng Kung University [K111-B094], [K111-B095], National Cheng Kung University Hospital, Taiwan [NCKUH-11204031], [NCKUMCS2022057].

1. Dyrking av C. glabrata
MERK: En multidrugresistent C. glabrata (C2-1000907) som er resistent mot de fleste antifungale midler, inkludert flukonazol, brukes til forsøkene. De eksperimentelle forholdene må kanskje tilpasses den spesifikke stammen, da variasjoner kan eksistere mellom forskjellige stammer. Alle forsøkene brukte log-fase Candida dyrket ved 25 ° C (etterligner naturlig infeksjon) for konsistens. C. glabratas mangel på hyfer forenkler...

Analyse av dannelse av små kolonier og medikamentresistens i Candida glabrata

<ol>
	<li>Soriano, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Invasive+candidiasis:+current+clinical+challenges+and+unmet+needs+in+adult+populations.">Invasive candidiasis: current clinical challenges and unmet needs in adult populations.</a> J Antimicrob Chemother. 78 (7), 1569-1585 (2023).</li><li>Pappas, P. G., Lionakis, M. S., Arendrup, M. C., Ostrosky-Zeichner, L., Kullberg, B. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Invasive+candidiasis.">Invasive candidiasis.</a> Nat Rev Dis Primers. 4, 18026 (2018).</li><li>Meyahnwi, D., Siraw, B. B., Reingold, A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Epidemiologic+features,+clinical+characteristics,+and+predictors+of+mortality+in+patients+with+candidemia+in+Alameda+County,+California;+a+2017-2020+retrospective+analysis.">Epidemiologic features, clinical characteristics, and predictors of mortality in patients with candidemia in Alameda County, California; a 2017-2020 retrospective analysis.</a> BMC Infect Dis. 22 (1), 843 (2022).</li><li>Whittaker, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+petite+mutation+in+yeast.">The petite mutation in yeast.</a> Subcell Biochem. 6, 175-232 (1979).</li><li>Hatab, M. A., Whittaker, P. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+respiration-deficient+mutants+from+the+pathogenic+yeast+Candida+albicans.">Isolation and characterization of respiration-deficient mutants from the pathogenic yeast Candida albicans.</a> Antonie Van Leeuwenhoek. 61 (3), 207-219 (1992).</li><li>Defontaine, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In-vitro+resistance+to+azoles+associated+with+mitochondrial+DNA+deficiency+in+Candida+glabrata.">In-vitro resistance to azoles associated with mitochondrial DNA deficiency in Candida glabrata.</a> J Med Microbiol. 48 (7), 663-670 (1999).</li><li>Brun, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Relationships+between+respiration+and+susceptibility+to+azole+antifungals+in+Candida+glabrata.">Relationships between respiration and susceptibility to azole antifungals in Candida glabrata.</a> Antimicrob Agents Chemother. 47 (3), 847-853 (2003).</li><li>Bouchara, J. P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=In-vivo+selection+of+an+azole-resistant+petite+mutant+of+Candida+glabrata.">In-vivo selection of an azole-resistant petite mutant of Candida glabrata.</a> J Med Microbiol. 49 (11), 977-984 (2000).</li><li>Badrane, H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genotypic+diversity+and+unrecognized+antifungal+resistance+among+populations+of+Candida+glabrata+from+positive+blood+cultures.">Genotypic diversity and unrecognized antifungal resistance among populations of Candida glabrata from positive blood cultures.</a> Nat Commun. 14 (1), 5918 (2023).</li><li>Shantal, C. -. J. N., Juan, C. -. C., Lizbeth, B. -. U. S., Carlos, H. -. G. J., Estela, G. -. P. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Candida+glabrata+is+a+successful+pathogen:+An+artist+manipulating+the+immune+response.">Candida glabrata is a successful pathogen: An artist manipulating the immune response.</a> Microbiol Res. 260, 127038 (2022).</li><li>Gamarra, S., Mancilla, E., Dudiuk, C., Garcia-Effron, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Candida+dubliniensis+and+Candida+albicans+differentiation+by+colony+morphotype+in+Sabouraud-triphenyltetrazolium+agar.">Candida dubliniensis and Candida albicans differentiation by colony morphotype in Sabouraud-triphenyltetrazolium agar.</a> Rev Iberoam Micol. 32 (2), 126-128 (2015).</li><li>Hung, J. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rose+bengal-mediated+photodynamic+therapy+to+inhibit+Candida+albicans.">Rose bengal-mediated photodynamic therapy to inhibit Candida albicans.</a> J Vis Exp. (181), e63558 (2022).</li><li>Cardoso, D. R., Franco, D. W., Olsen, K., Andersen, M. L., Skibsted, L. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Reactivity+of+bovine+whey+proteins,+peptides,+and+amino+acids+toward+triplet+riboflavin+as+studied+by+laser+flash+photolysis.">Reactivity of bovine whey proteins, peptides, and amino acids toward triplet riboflavin as studied by laser flash photolysis.</a> J Agric Food Chem. 52 (21), 6602-6606 (2004).</li><li>Hall, R. M., Trembath, M. K., Linnane, A. W., Wheelis, L., Criddle, R. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Factors+affecting+petite+induction+and+the+recovery+of+respiratory+competence+in+yeast+cells+exposed+to+ethidium+bromide.">Factors affecting petite induction and the recovery of respiratory competence in yeast cells exposed to ethidium bromide.</a> Mol Gen Genet. 144 (3), 253-262 (1976).</li><li>Chen, X. J., Clark-Walker, G. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+petite+mutation+in+yeasts:+50+years+on.">The petite mutation in yeasts: 50 years on.</a> Int Rev Cytol. 194, 197-238 (2000).</li><li>Piskur, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inheritance+of+the+yeast+mitochondrial+genome.">Inheritance of the yeast mitochondrial genome.</a> Plasmid. 31 (3), 229-241 (1994).</li><li>Wong, T. W., Wang, Y. Y., Sheu, H. M., Chuang, Y. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Bactericidal+effects+of+toluidine+blue-mediated+photodynamic+action+on+Vibrio+vulnificus.">Bactericidal effects of toluidine blue-mediated photodynamic action on Vibrio vulnificus.</a> Antimicrob Agents Chemother. 49 (3), 895-902 (2005).</li><li>Wong, T. W., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Indocyanine+green-mediated+photodynamic+therapy+reduces+methicillin-resistant+Staphylococcus+aureus+drug+resistance.">Indocyanine green-mediated photodynamic therapy reduces methicillin-resistant Staphylococcus aureus drug resistance.</a> J Clin Med. 8 (3), 411 (2019).</li><li>Warrier, A., Mazumder, N., Prabhu, S., Satyamoorthy, K., Murali, T. S. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Photodynamic+therapy+to+control+microbial+biofilms.">Photodynamic therapy to control microbial biofilms.</a> Photodiagnosis Photodyn Ther. 33, 102090 (2021).</li><li>Hung, J. H., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+photodynamic+therapy+against+fungal+keratitis.">Recent advances in photodynamic therapy against fungal keratitis.</a> Pharmaceutics. 13 (12), 2011 (2021).</li></ol>

Denne studien avslører PDT som den første rapporterte metoden for å indusere petite kolonidannelse i Candida, som overgår de etablerte effektene av ethidiumbromid og flukonazol. Denne nye observasjonen nødvendiggjør videre utforskning for å avdekke implikasjonene for både sopputryddelse ved å redusere virulens og fremveksten av resistensmekanismer.
RB-mediert PDT hemmer effektivt veksten av C. glabrata, noe som tyder på en potensiell alternativ behandlingstilnærming for Can...

Soppinfeksjoner, spesielt de som er forårsaket av Candida albicans og stadig mer stoffresistente Candida glabrata, utgjør en alvorlig global trussel1. Disse infeksjonene kan være dødelige, spesielt for pasienter på sykehus og de med svekket immunforsvar. Økende soppdrepende resistens truer kontrollen av invasiv candidiasis, en alvorlig soppinfeksjon med høy dødelighet, spesielt fra Candida albicans2. Resistente stammer hindrer effektiv behandling, noe som potensielt øker både kompleksiteten og dødsraten. I Alameda County, California, USA, har C. glabrata blitt den mest utbredte invasive arten3. Dette skiftet i utbredelsen og distribusjonen av Candida-arter kan påvirkes av lokal helsepraksis, pasientdemografi, bruk av antifungale midler og forekomsten av risikofaktorer for Candida-infeksjoner.
Petite mutanter i Candida, som mangler funksjonelle mitokondrier, avslører hvordan denne organellen påvirker legemiddelrespons, virulens og stressmotstand 4,5. C. glabrata danner lett disse koloniene, og får følsomhet for polyener mens de mister den til azoler6. Azolfølsomhet og respirasjonsfunksjon er intrikat forbundet, med redusert respirasjon som fører til resistens via mitokondrielt DNA-tap7. Petitekolonier av C. glabrata med azolresistens er isolert fra avføringsprøver fra benmargstransplanterte som gjennomgår flukonazolbehandling8 og fra blodkulturflasker hos pasienter med blodbaneinfeksjoner9. Deres potensielle implikasjoner i stoffresistens, virulens og stressrespons fremhever deres kliniske betydning. I tillegg gjør deres distinkte egenskaper dem til verdifulle verktøy for å undersøke grunnleggende spørsmål i mitokondriebiologi5. Etter hvert som forskningen på små mutanter fortsetter, vil deres anvendelser i både klinisk og grunnleggende forskning sannsynligvis utvides.
Denne studien oppdaget at fotodynamisk terapi (PDT) kan indusere petite kolonier i C. glabrata, og utvide rekkevidden av metoder utover de tradisjonelle teknikkene for å eksponere C. glabrata for ethidiumbromid eller flukonazol.

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.22 &#956;m filter</td>
			<td>Merck, Taipei, Taiwan</td>
			<td>Millex, SLGVR33RS</td>
			<td></td>
		</tr>
		<tr>
			<td>1.5 mL microfuge tube</td>
			<td>Neptune, San Diego, USA</td>
			<td>#3745</td>
			<td></td>
		</tr>
		<tr>
			<td>20% Triphenyltetrazolium chloride (TTC)</td>
			<td>Sigma-Aldrich, MO, USA</td>
			<td>T8877</td>
			<td></td>
		</tr>
		<tr>
			<td>5 mL polypropylene round bottom tube</td>
			<td>Corning, AZ, USA</td>
			<td>352059</td>
			<td></td>
		</tr>
		<tr>
			<td>5 mL round-bottom tube with cell strainer cap</td>
			<td>Corning, AZ, USA</td>
			<td>Falcon, #352235</td>
			<td></td>
		</tr>
		<tr>
			<td>96-well plate</td>
			<td>Alpha plus, Taoyuan Hsien, Taiwan</td>
			<td>#16196</td>
			<td></td>
		</tr>
		<tr>
			<td>Agar</td>
			<td>BRS, Tainan, Taiwan</td>
			<td>AG012</td>
			<td></td>
		</tr>
		<tr>
			<td>Blank disk</td>
			<td>Advantec, Tokyo, Japan</td>
			<td>49005040</td>
			<td></td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Eppendorf, UK</td>
			<td>5415R</td>
			<td></td>
		</tr>
		<tr>
			<td>Ethidium bromide solution</td>
			<td>Sigma-Aldrich, MO, USA</td>
			<td>E1510</td>
			<td></td>
		</tr>
		<tr>
			<td>Fluconazole, 2 mg/mL</td>
			<td>Pfizer, NY, USA</td>
			<td>BC18790248</td>
			<td></td>
		</tr>
		<tr>
			<td>GraphPad Prism</td>
			<td>GraphPad Software</td>
			<td>Version 7.0</td>
			<td></td>
		</tr>
		<tr>
			<td>Green light emitting diode (LED) strip</td>
			<td>Nanyi electronics Co.,Ltd, Tainan, Taiwan</td>
			<td>5050</td>
			<td>Excitation wave: 500~550 nm</td>
		</tr>
		<tr>
			<td>Low Temperature. shake Incubators</td>
			<td>Yihder, Taipei, Taiwan</td>
			<td>LM-570D (R)</td>
			<td></td>
		</tr>
		<tr>
			<td>Mouth care cotton swabs</td>
			<td>Good Verita Enterprise, Taipei, Taiwan</td>
			<td>161357</td>
			<td></td>
		</tr>
		<tr>
			<td>Muller Hinton II agar</td>
			<td>BD biosciences, California, USA</td>
			<td>211438</td>
			<td></td>
		</tr>
		<tr>
			<td>Multimode microplate reader</td>
			<td>Molecular Devices</td>
			<td>SpectraMax i3x</td>
			<td></td>
		</tr>
		<tr>
			<td>OD600 spectrophotometer</td>
			<td>Biochrom, London, UK</td>
			<td>Ultrospec 10</td>
			<td></td>
		</tr>
		<tr>
			<td>Rose Bengal</td>
			<td>Sigma-Aldrich, USA</td>
			<td>330000</td>
			<td>stock concentration 40 mg/mL = 4%, prepare in PBS, stored at 4 &#176;C</td>
		</tr>
		<tr>
			<td>Sterilized glass tube</td>
			<td>Sunmei, Tainan, Taiwan</td>
			<td>AK45048-16100</td>
			<td></td>
		</tr>
		<tr>
			<td>Yeast Extract Peptone Dextrose Medium</td>
			<td>HIMEDIA, India</td>
			<td>M1363</td>
			<td></td>
		</tr>
	</tbody>
</table>

induction of petite colonies in candida glabrate via rose bengal-mediated photodynamic therapy

Overfor en dødelighet på 40% hos candidemi-pasienter, er stoffresistent Candida og deres petite mutanter fortsatt en stor behandlingsutfordring. Antimikrobiell fotodynamisk terapi (aPDT) retter seg mot flere soppstrukturer, i motsetning til antibiotika / antifungale midler, som potensielt hindrer resistens. Tradisjonelle metoder for å indusere petite kolonier stole på ethidiumbromid eller flukonazol, noe som kan påvirke stofffølsomhet og stressresponser. Denne studien undersøkte anvendelsen av grønt lys (topp 520 nm) og rose bengal (RB) fotosensibilisator for å bekjempe et stoffresistent Candida glabrata-isolat . Funnene viste at aPDT-behandling signifikant hemmet cellevekst (≥99,9% reduksjon) og effektivt induserte petite kolonidannelse, som det fremgår av redusert størrelse og tap av mitokondriell redoksindikatorfarging. Denne studien gir innledende bevis på at aPDT kan indusere petite kolonier i en multidrugresistent C. glabrata-stamme in vitro, og tilbyr en potensielt transformativ tilnærming for å bekjempe resistente soppinfeksjoner.

Fungal infections, particularly those caused by Candida albicans and increasingly drug-resistant Candida glabrata, pose a serious global threat1. These infections can be deadly, especially for hospitalized patients and those with weakened immune systems. Rising antifungal resistance threatens the control of invasive candidiasis, a severe fungal infection with high mortality, especially from Candida albicans2. Resistant strains hinder effective treatment, potentially increasing both complexity and death rates. In Alameda County, California, USA, C. glabrata has become the most prevalent invasive species3. This shift in the prevalence and distribution of Candida species may be influenced by local healthcare practices, patient demographics, the utilization of antifungal agents, and the prevalence of risk factors for Candida infections.
Petite mutants in Candida, lacking functional mitochondria, reveal how this organelle affects drug response, virulence, and stress resistance4,5. C. glabrata readily forms these colonies, gaining sensitivity to polyenes while losing it to azoles6. Azole sensitivity and respiratory function are intricately linked, with diminished respiration leading to resistance via mitochondrial DNA loss7. Petite colonies of C. glabrata with azole resistance have been isolated from human stool samples from a bone marrow transplant recipient undergoing fluconazole treatment8 and from blood culture bottles of patients with bloodstream infections9. Their potential implications in drug resistance, virulence, and stress response highlight their clinical significance. Additionally, their distinct properties make them valuable tools for investigating fundamental questions in mitochondrial biology5. As research into petite mutants continues, their applications in both clinical and basic research are likely to expand.
This study discovered that photodynamic therapy (PDT) can induce petite colonies in C. glabrata, expanding the range of methods beyond the traditional techniques of exposing C. glabrata to ethidium bromide or fluconazole.

Forfatter Spotlight: Fotodynamisk terapi som en ny tilnærming for å indusere små kolonier i medikamentresistent Candida for soppdrepende forskning

Induksjon av petite kolonier i Candida glabrate via Rose Bengal-mediert fotodynamisk terapi

The research aims to use photodynamic therapy, also known as PDT, to fight against drug-resistant Candida infection. In this study, we are trying to answer how effectively PDT kills the fungi, and how the virulence of the fungi changes after the treatment. We found petite colonies from the after PDT.Present methods for inducing petite colonies rely on ethidium bromide or fluconazole, which may work through different pathways on drug resistance and the stress responses, compared to PDT. Recently, petite colonies of Candida have become a significant concern in the clinic because they may resist medication and increase mortality rates. Photodynamic therapy has become one of the antifungal treatments and our finding helps breach the gap between traditional methods used to study petite colonies.Our findings provide evidence that PDT can induce petite colonies in drug resistant Candida in-vitro, presenting a groundbreaking method to study petite colonies. We'll definitely delve deeper into this field. There are many questions to be answered.How and why do petite colonies form after PDT stress? Do different Candida species respond differently? I believe it's a never ending story to be explored.

Dataene presenteres som gjennomsnittet med ± standardfeil og ble hentet fra tre uavhengige eksperimenter, med minst triplikater i hver gruppe. Eksperimentelle data, inkludert kolonitelling, OD600-målinger og TTC-fargeresultater, ble grafet og statistisk analysert ved hjelp av grafisk og statistisk programvare (se materialfortegnelse). Enveis ANOVA eller t-test ble brukt for å analysere dataene, og en p-verdi <0,05 ble vurdert som signifikant. Skanning ble utført med en 48...

Watch this Scientific Journal Video about Photodynamic Therapy as a Novel Approach to Induce Petite Colonies in Drug-Resistant Candida for Antifungal Research at JoVE.com

Betydningen av små kolonier i Candida spp. stoffresistens har ikke blitt fullstendig utforsket. Antimikrobiell fotodynamisk terapi (aPDT) tilbyr en lovende strategi mot stoffresistente soppinfeksjoner. Denne studien viser at rose bengal-mediert aPDT effektivt deaktiverer Candida glabrata og induserer petite kolonier, og presenterer en unik prosedyre.

Facing a 40% mortality rate in candidemia patients, drug-resistant Candida and their petite mutants remain a major treatment challenge. Antimicrobial ...

Forskningen tar sikte på å bruke fotodynamisk terapi, også kjent som PDT, for å bekjempe stoffresistent Candida-infeksjon. I denne studien prøver vi å svare på hvor effektivt PDT dreper soppene, og hvordan soppens virulens endres etter behandlingen. Vi fant små kolonier fra etter PDT.Nåværende metoder for å indusere petite kolonier stole på ethidiumbromid eller flukonazol, som kan virke gjennom forskjellige veier på stoffresistens og stressresponsene, sammenlignet med PDT. Nylig har små kolonier av Candida blitt en betydelig bekymring i klinikken fordi de kan motstå medisinering og øke dødeligheten. Fotodynamisk terapi har blitt en av de antifungale behandlingene, og vårt funn bidrar til å bryte gapet mellom tradisjonelle metoder som brukes til å studere petite kolonier.Våre funn gir bevis på at PDT kan indusere petite kolonier i stoffresistent Candida in-vitro, og presenterer en banebrytende metode for å studere petite kolonier. Vi vil definitivt dykke dypere inn i dette feltet. Det er mange spørsmål som skal besvares.Hvordan og hvorfor dannes små kolonier etter PDT-stress? Reagerer forskjellige Candida-arter forskjellig? Jeg tror det er en uendelig historie som skal utforskes.

induction-petite-colonies-candida-glabrate-via-rose-bengal-mediated

Research

JoVE Journal

Biology

Induction of Petite Colonies in Candida glabrate via Rose Bengal-Mediated Photodynamic Therapy

Facing a 40% mortality rate in candidemia patients, drug-resistant Candida and their petite mutants remain a major treatment challenge. Antimicrobial photodynamic therapy (aPDT) targets multiple fungal structures, unlike antibiotics/antifungals, potentially thwarting resistance. Traditional methods for inducing petite colonies rely on ethidium bromide or fluconazole, which can influence drug susceptibility and stress responses. This study investigated the application of green light (peak 520 nm) and rose bengal (RB) photosensitizer to combat a drug-resistant Candida glabrata isolate. The findings revealed that aPDT treatment significantly inhibited cell growth (&#8805;99.9% reduction) and effectively induced petite colony formation, as evidenced by reduced size and loss of mitochondrial redox indicator staining. This study provides initial evidence that aPDT can induce petite colonies in a multidrug-resistant C. glabrata strain in vitro, offering a potentially transformative approach for combating resistant fungal infections.

The significance of petite colonies in Candida spp. drug resistance has not been fully explored. Antimicrobial photodynamic therapy (aPDT) offers a promising strategy against drug-resistant fungal infections. This study demonstrates that rose bengal-mediated aPDT effectively deactivates Candida glabrata and induces petite colonies, presenting a unique procedure.