In Vivo Efficacy of Amphotericin B against Four Candida auris Clades
<p>Survival of neutropenic AMB-treated and control BALB/c mice infected with the isolates from the four <span class="html-italic">Candida auris</span> clades: isolates 27 and 196 (<b>A</b>) (South Asian clade), isolates CBS 12,372 {12,372} and CBS 12,373 {12,373} (<b>B</b>) (East Asian clade), isolates 2 and 204 (<b>C</b>) (South African clade) and isolates I-24 and I-156 (<b>D</b>) (South American (clade). Two environmental isolates, CDC B-13,108 {13,108} and CDC B-13,112 {13,112} (<b>E</b>), from the South American clade were also tested. The infectious dose was 10<sup>7</sup> CFU/mouse. Daily 1 mg/kg of amphotericin B (AMB) treatment began 24 h postinfection. After 21 days the survival rate was analyzed by Kaplan–Meier test. Blue ellipses in survival curves in cases of mice infected with isolates 12,372 and 12,373 (East Asian clade) (<b>B</b>), isolate 2 (South African clade) (<b>C</b>) and isolate I-156 (South American clade) (<b>E</b>) indicate that these mice showed ataxia 2–3 days before their death.</p> "> Figure 2
<p>Fungal kidneys, heart and brain burdens were determined with 2-2 clinical isolates from the South Asian {isolates 27 (<b>A</b>) and 196 (<b>B</b>)}, East Asian {isolates CBS 12,372 (<b>C</b>) and CBS 12,373 (<b>D</b>)}, South African {isolates 2 (<b>E</b>) and 204 (<b>F</b>)} and South American {isolates I-24 (<b>G</b>) and I-156 (<b>H</b>)} clades at day 6. Two environmental isolates CDC B-13108 (<b>I</b>) and CDC B-13112 (<b>J</b>) from the South American clade were also tested. The infectious dose was 8 × 10<sup>6</sup> CFU/mouse. Daily 1 mg/kg of amphotericin B (AMB) treatment began 24 h post-infection. The bars represent the medians. Level of statistical significance is indicated at <span class="html-italic">p</span> < 0.05 (*), <span class="html-italic">p</span> < 0.01 (**). NS corresponds to non-significant.</p> "> Figure 3
<p>The effect of 2 days amphotericin B (AMB) treatment on the CFU in the urine in mice infected with isolates 196 (South Asian clade), 12,372 (East Asian clade), 204 (South African clade), I-156 (bloodstream isolate from the South American clade) and 13,112 (environmental isolate from the South American clade). Urine was collected on day 3. The bars represent the medians. Level of statistical significance is indicated at <span class="html-italic">p</span> < 0.001.</p> "> Figure 4
<p>Histopathological examination of the heart, kidney and brain with Periodic Acid Schiff or Mallory’s PTAH staining from amphotericin B (AMB) treated mice found dead after intravenously challenged with <span class="html-italic">Candida auris</span> isolates. In mice infected with isolate 196 (South Asian clade) which died on day 5, kidneys (<b>A</b>) showed large multifocal fungal lesions, fungal cells were detected in the parenchyma, necrosis occurred in the tubular tissue, but glomeruli were not invaded. In the heart (<b>B</b>), <span class="html-italic">C. auris</span> produced large aggregates and coagulative necrosis of myocytes. Mallory’s PTAH staining always revealed contraction band necrosis (myofibrillar degeneration) (<b>C</b>). In the case of mice infected with isolate 12,372 (East Asian clade) which died on day 6, in the hearts multifocal (<b>D</b>) or only focal (<b>E</b>) fungal infiltrates were found. In the kidneys (<b>F</b>) small lesions were detectable (black arrow). In mice infected with isolate 204 (South African clade) which died on day 5, 4–5 aggregates were seen in the hearts (<b>G</b>) with contraction band necrosis (<b>H</b>). In the cerebellum (<b>I</b>), one large and one smaller PAS positive fungal lesions were seen. In mice infected with isolate I-156 (South American clade) which died on day 5, multifocal, large aggregates were seen in the hearts (<b>J</b>) and kidneys (<b>K</b>) with abundant yeast cells in the cerebrum (<b>L</b>). In mice infected with isolate CDC B-13108 which died on day 4, the heart showed large and small multifocal infiltrates (<b>M</b>) with contraction band necrosis (<b>N</b>). In the cerebrum (<b>O</b>), yeast cells were visible. Fungal lesions were indicated with black arrows. Magnification, (<b>A</b>,<b>B</b>,<b>D</b>–<b>G</b>,<b>J</b>,<b>K,M</b>) ×100, (<b>C</b>,<b>H</b>,<b>I</b>,<b>L</b>,<b>N,O</b>) ×400.</p> "> Figure 5
<p>Histopathological examination of the cerebrum, cerebellum and kidney with Periodic Acid Schiff staining from a neutropenic, amphotericin B treated, mice infected with isolate 12,372 (East Asian clade) which died on day 20. The cerebrum (<b>A</b>) and cerebellum (<b>B</b>) showed yeast cells, but kidneys (<b>C</b>) were sterile, with normal histology without fungal cells. Fungal lesions were indicated with black arrows. Magnification, ×100.</p> "> Figure 6
<p>Histopathological findings of the heart, kidney and cerebrum with Periodic Acid Schiff staining from fungal burden experiments. Neutropenic mice infected with isolate I-156 (South American clade) sacrificed and dissected on day 2 showed small fungal foci in the hearts (<b>A</b>) and kidneys (<b>B</b>), but the cerebrum (<b>C</b>) did not contain fungal cells. The histopathology regardless that mice were AMB-treated or not was the same for all isolates and clades on day 2. In case of a mouse infected with isolate 12,372 (East Asian clade) and treated with 1 mg/kg of AMB, on day 6 the heart showed normal histology without fungal involvement (<b>D</b>), but the kidney (<b>E</b>) and cerebrum (<b>F</b>) showed fungal cells. Fungal lesions were indicated with black arrows. Magnification, ×100.</p> ">
Abstract
:1. Introduction
2. Materials and Methods
2.1. Isolates
Mice and Immunosuppression
2.2. Lethality Experiments
2.3. Fungal Tissue Burden Experiments
2.4. Urine Collection
2.5. Histopathology
3. Results
3.1. Lethality Experiments
3.1.1. South Asian Clade
3.1.2. East Asian Clade
3.1.3. South African Clade
3.1.4. South American Clade
3.2. Fungal Burden Experiments
3.2.1. South Asian Clade
3.2.2. East Asian Clade
3.2.3. South African Clade
3.2.4. South American Clade
3.3. Urine Culture
3.4. Histopathology
3.4.1. Lethality Experiments
South Asian Clade
East Asian Clade
South African Clade
South American Clade
3.4.2. Fungal Burden Experiments
4. Discussion
5. Conclusions
Supplementary Materials
Author Contributions
Funding
Institutional Review Board Statement
Informed Consent Statement
Data Availability Statement
Conflicts of Interest
References
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Clade | Isolate Number | Body Site |
---|---|---|
South Asian | 27 (NCPF 89891) | Pleural fluid |
196 | Blood | |
East Asian | 12372 (CBS 12372) | Blood |
12373 (CBS 12373) | Blood | |
South African | 2 (NCPF 8977) | Cerebrospinal fluid |
204 | Tracheostomy | |
South American | 13108 (CDC B-13108) | Hospital environment |
13112 (CDC B-13112) | Hospital environment | |
I-24 | Blood | |
I-156 | Blood |
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Forgács, L.; Borman, A.M.; Kovács, R.; Balázsi, D.; Tóth, Z.; Balázs, B.; Chun-Ju, C.; Kardos, G.; Kovacs, I.; Majoros, L. In Vivo Efficacy of Amphotericin B against Four Candida auris Clades. J. Fungi 2022, 8, 499. https://doi.org/10.3390/jof8050499
Forgács L, Borman AM, Kovács R, Balázsi D, Tóth Z, Balázs B, Chun-Ju C, Kardos G, Kovacs I, Majoros L. In Vivo Efficacy of Amphotericin B against Four Candida auris Clades. Journal of Fungi. 2022; 8(5):499. https://doi.org/10.3390/jof8050499
Chicago/Turabian StyleForgács, Lajos, Andrew M. Borman, Renátó Kovács, Dávid Balázsi, Zoltán Tóth, Bence Balázs, Chiu Chun-Ju, Gábor Kardos, Ilona Kovacs, and László Majoros. 2022. "In Vivo Efficacy of Amphotericin B against Four Candida auris Clades" Journal of Fungi 8, no. 5: 499. https://doi.org/10.3390/jof8050499