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Файл:Ординатура / Хирургия / Библиотека им академика М.И. Перельмана / Книга_5219_Библиотеки_им_академика_М_И_Перельмана.pdf
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G. Gangwar et al.
monitoring of biolm biomass provided additional conrmation of the results.
Biolm formation was signicantly reduced in groups treated with torin2, and inbetween erosions were observed. Similarly, biolm formation was less dense in the
rapamycin-treated group than in the vehicle control group, and there was visible
biolm erosion in several locations. Comparing the biolm biomass to the corresponding vehicle controls, TOR inhibition reduced it by approximately tenfold. It
has previously been demonstrated that TOR plays a signicant role in controlling
the expression of adhesion and efux genes in Candida albicans (Kumar etal.
2018). The starvation responses and biolm formation of Candida albicans are
regulated by the TOR-activating GTPases Gtr1 and Rhb1 (Flanagan etal. 2017).
However, this study provided evidence that, even in C. auris, TOR inhibition
reduces the formation of biolms. Nevertheless, the function of TOR in the expansion of biolm cells was substantiated by in-depth microscopy of biolms exposed
to Tor inhibitors. This was the rst report demonstrating the signicant reduction in
biolm growth invitro following TOR inhibition with torin2 and rapamycin (Biswas
etal. 2023a, b; Kumar etal. 2018).
10.14 Mixed Biofilm inC. auris
In nature biolms are produced by a variety of microbial species, and these mixedspecies biolms accurately reect the living conditions of fungi, bacteria, viruses
(phages), and/or protozoa. Microorganisms that collaborate and compete can be
found in mixed-species biolms. The biolms of mixed species in C. auris have not
been thoroughly investigated. However, since C. auris colonizes skin that has its
own microora, there is debate regarding the formation of mixed-species biolm.
Research has been done to comprehend the mixed-species biolm of Staphylococcus
and C. auris. Although the non-aggregating C. auris and Staphylococcus sp. inter-
action does not affect the biolm mass, it is evident that bacteria contribute to the
environmental persistence of the C. auris strain. Since mixed-species biolm can
proliferate at 37°C just like bacteria, it is important to investigate the fundamentals
of this relationship with skin microbiota (Khari etal. 2023).
Along with a variety of bacterial microbiomes, C. auris can colonize human skin
over time. Study on the detrimental effects of antiseptics on Staphylococci-
containing dual-species interkingdom biolms revealed that for an exposure at 2%,
10%, and 3%, respectively, biolm viable cell counts were signicantly reduced by
hydrogen peroxide (H2O2), povidone iodine, and chlorhexidine. Remarkably, biolms treated with H2O2 demonstrated a substantial ability to recuperate and greatly
proliferate after therapy. Fortunately, the resistance of C. auris against antiseptics
invitro was not increased by inter-kingdom interactions in dual-species biolms of
C. auris and staphylococci. These data show that mixed infections can be treated
with povidone iodine and chlorhexidine, but H2O2 should be used with caution
(Gülmez etal. 2022).
10 Dissemination ofCandida auris Biolms: AMedical Abrosia
273
10.15 Quantification ofBiofilm
Different laboratories use different methods for the quantication of biolms. In
routine, an aseptic, at-bottomed, 96-well microplate is inseminated with 100μL of
the standard test organism inoculum per well in order to form single or mixed biolms. Then the plate is stored in an incubator for 24h at 37°C.Measure total biolm mass using a crystal violet (CV) staining scheme and take absorbance at 570nm
with a microtiter plate reader. According to the manufacturer’s instructions, biolms
vital biomass is quantied by using the tetrazolium 2,3-bis(2-methoxy-4-nitro-5
sulfophenyl)-5-[(phenylamine) carbonyl]-2H-hydroxide reduction assay (XTT)
(Sigma-Aldrich, St. Louis, MO, USA). With the help of a microtiter plate reader, the
absorbance of the resultant solution is determined at 492nm. To further understand
the formation of mixed biolms, the CFU assay was used. Adhered biolms are, in
short, thoroughly scraped and serially diluted in PBS (phosphate-buffered saline).
On TSA agar plates supplemented with amphotericin B (for K. pneumoniae) and
Rose Bengal Agar plates supplemented with chloramphenicol (for C. auris), the
diluted suspension of cells is spread. After incubating the biolm cells for 24h at
37°C, the CFU count of the biolm cells is determined. The viability of each assay
is assessed using mean log CFU (Maione etal. 2022).
10.16 Antifungal Resistance Is aCharacteristic Feature
ofCandida Biofilms
Biolm formation is responsible for increasing the resistance to antifungals. It is
one of the main factors responsible for drug tolerance. Extracellular matrix of
C. auris biolm is made up of mannan and glucan that sequester drugs to prevent
binding with their target. This antifungal sequestration provides resistance to various drugs (Cernakova etal. 2021).
There are mainly three classes of drugs such as azoles, polyenes, and echinocan-
dins. Lanosterol-1,4-alpha-demethylase (LD) is the target of azoles. This protein
helps in the conversion of lanosterol to ergosterol. The target of echinocandin is
β-1,3--glucan synthase, a component of the fungal cell wall. Ergosterol is the target of polyenes that affects membrane permeability by forming pores. While the
target of nucleoside analogue is thymidine synthase and it inhibits DNA and RNA
synthesis (Chaabane etal. 2019).
Mutation in some genes tends to be responsible for resistance to these drugs.
Mutations in ERG11 and TAC1b (some mutations Y132F, K143R, and F126L)
genes are responsible for the resistance to the azoles. A large number of uconazoleresistant strains have mutations in the TAC1b gene (Li etal. 2021b; Rhodes etal.
2018). The tolerance of C. auris to ucytosine is due to F211I amino acid substitu-
tion in the FUR1 gene (Gade etal. 2020). Resistance to echinocandin is rare and it
occurs due to the mutation at single amino acid S639in the FKS1 gene (Vandeputte
etal. 2008). While the resistance to polyenes is not still clear recent studies have
reported that resistance to polyenes is associated with the mutation in genes
274
responsible for ergosterol biosynthesis (Hull etal. 2012; Escandon et al. 2019).
Genes involved in polyene resistance are ERG1, ERG2, ERG3, ERG5, ERG6,
ERG11, and ERG13 (Chybowska etal. 2020; Ciurea etal. 2021).
G. Gangwar et al.
10.17 Therapeutic Approaches toReduce C. auris Biofilm
A variety of therapeutic strategies have been explored to eradicate C. auris biolm
such as photodynamic therapy (Tan etal. 2019), SCY-078 (Jallow and Govender
2021), NFAP2 (Kovács et al. 2021), bismuth nanoparticles (BiNPs) (Vazquez-
Munoz etal. 2020), silver nanoparticle (AgNPs) (Lara etal. 2020; AlJindan and
AlEraky 2022), 6-shogaol (Kim and Eom 2021), nitric oxide (NO) nanoparticles
(Cleare etal. 2020), antifungal peptide derivative Cm-p5 (Kubiczek etal. 2020),
minocycline-EDTA-ethanol antimicrobial catheter lock solution (Reitzel et al.
2020), Lavandula angustifolia essential oil (de Alteriis etal. 2021), and defensin-
like protein 1 (D-lp1) (Kamli etal. 2022). Some of them are discussed below.
10.18 Photodynamic Therapy
PD therapy is a two-step process that works on the interaction of light energy with
photosensitive compounds and molecular oxygen. This causes ROS species to
develop, which in turn causes cell death. There are three regions in the visible light
spectrum, every light region contributes differently to the inhibition of the C. auris
biolm. Data from earlier studies indicate all light plus PS compounds have an
effective role in inhibiting biolm formation during the developmental and dispersion phases of biolm formation, none of them are effective at inhibiting biolm
formation during the adherence stage (Tan etal. 2019).
10.19 Impact ofSCY-078: ANovel Inhibitor ontheFormation
ofBiofilms
SCY-078 or IbrexafungerP (IBX) is the rst oral antifungal medication available. It
inhibits the formation of fungal cell wall as it acts as the non-competitive inhibitor
of 1,3-β--glucan synthase. IBX functions similar to the echinocandins but their
binding sites are not identical. Like echinocandins, it has a fungicidal effect on
Candida species and fungistatic effects on Aspergillus species (Jallow and
Govender 2021).
10 Dissemination ofCandida auris Biolms: AMedical Abrosia
275
10.20 Effect ofNeosartorya fischeri Antifungal Protein
2 (NFAP2)
Neosartorya scheri antifungal protein 2 (NFAP2) is a cationic, cysteine-containing
protein isolated from lamentous ascomycetes. NFAP2 permeabilizes cell membrane resulting in osmotic stress and ultimately leads to cell death (Kovács
etal. 2021).
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G. Gangwar et al.
Polymicrobial Biofilms ofC. albicans
withBacterial Species: AnInsight into
Intergenus Interaction
PurviJoshi, PinalTrivedi, RohitBhattacharjee,
MuskanSahu, andDevarshiGajjar
Abstract
Polymicrobial biolms are more prevalent than reported in the clinical scenario
and are more complex and harmful to the host. C. albicans, being the most prevalent Candida species causing infections, has been found to colonise and infect
immunocompromised humans. C. albicans is found to interact with various bacterial species like Streptococcus, Staphylococcus, Pseudomonas, and E. coli.
This chapter focuses on this intergenus interactions and associated antimicrobial
resistance.
Keywords
Polymicrobial biolms · Microbial interactions · Antimicrobial resistance ·
Candida-bacteria biolms
11
11.1 Introduction
Humans are colonised by various population of bacteria and fungi, forming a diverse
microbial community. These microorganisms can either positively or negatively
inuence the host and their interactions play a signicant role in determining the
overall well-being of the individual.
Candida species are the leading culprits behind fungal infections on a global
scale and are widely distributed within the human microbiota. Candida albicans, a
fungus that typically coexists with humans, can be found naturally on mucosal
P. Joshi · P. Trivedi · R. Bhattacharjee · M. Sahu · D. Gajjar (*)
Department of Microbiology and Biotechnology Centre, Faculty of Science, The Maharaja
Sayajirao University of Baroda, Vadodara, Gujarat, India
e-mail: devarshi.gajjar-microbio@msubaroda.ac.in
© The Author(s), under exclusive license to Springer Nature Singapore Pte
Ltd. 2024
S. Hameed, P. Vijayaraghavan (eds.), Recent Advances in Human Fungal
Diseases, https://doi.org/10.1007/978-981-97-4909-6_11
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