The Autofluorescence Patterns of <i>Acanthamoeba castellanii, Pseudomonas aeruginosa</i> and <i>Staphylococcus aureus</i>: Effects of Antibiotics and Tetracaine
<i>Acanthamoeba</i> Keratitis (AK) can lead to substantial vision loss and morbidity among contact lens wearers. Misdiagnosis or delayed diagnosis is a major factor contributing to poor outcomes of AK. This study aimed to assess the effect of two antibiotics and one anaesthetic drug used...
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| Language: | English |
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MDPI AG
2021-07-01
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| Series: | Pathogens |
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| Online Access: | https://www.mdpi.com/2076-0817/10/7/894 |
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| author | Hari Kumar Peguda Saabah B. Mahbub Tashi Doma Sherpa Dinesh Subedi Abbas Habibalahi Ayad G. Anwer Zi Gu Mark D. P. Willcox Ewa M. Goldys Nicole A. Carnt |
| author_facet | Hari Kumar Peguda Saabah B. Mahbub Tashi Doma Sherpa Dinesh Subedi Abbas Habibalahi Ayad G. Anwer Zi Gu Mark D. P. Willcox Ewa M. Goldys Nicole A. Carnt |
| author_sort | Hari Kumar Peguda |
| collection | DOAJ |
| description | <i>Acanthamoeba</i> Keratitis (AK) can lead to substantial vision loss and morbidity among contact lens wearers. Misdiagnosis or delayed diagnosis is a major factor contributing to poor outcomes of AK. This study aimed to assess the effect of two antibiotics and one anaesthetic drug used in the diagnosis and nonspecific management of keratitis on the autofluorescence patterns of <i>Acanthamoeba</i> and two common bacteria that may also cause keratitis. <i>Acanthamoeba castellanii</i> ATCC 30868, <i>Pseudomonas aeruginosa</i> ATCC 9027, and <i>Staphylococcus aureus</i> ATCC 6538 were grown then diluted in either PBS (bacteria) or ¼ strength Ringer’s solution (<i>Acanthamoeba</i>) to give final concentrations of 0.1 OD at 660 nm or 10<sup>4</sup> cells/mL. Cells were then treated with ciprofloxacin, tetracycline, tetracaine, or no treatment (naïve). Excitation–emission matrices (EEMs) were collected for each sample with excitation at 270–500 nm with increments in 5 nm steps and emission at 280–700 nm at 2 nm steps using a Fluoromax-4 spectrometer. The data were analysed using MATLAB software to produce smoothed color-coded images of the samples tested. <i>Acanthamoeba</i> exhibited a distinctive fluorescence pattern compared to bacteria. The addition of antibiotics and anaesthetic had variable effects on autofluorescence. Tetracaine altered the fluorescence of all three microorganisms, whereas tetracycline did not show any effect on the fluorescence. Ciprofloxacin produced changes to the fluorescence pattern for the bacteria, but not <i>Acanthamoeba</i>. Fluorescence spectroscopy was able to differentiate <i>Acanthamoeba</i> from <i>P. aeruginosa and S. aureus</i> in vitro. There is a need for further assessment of the fluorescence pattern for different strains of <i>Acanthamoeba</i> and bacteria. Additionally, analysis of the effects of anti-amoebic drugs on the fluorescence pattern of <i>Acanthamoeba</i> and bacteria would be prudent before in vivo testing of the fluorescence diagnostic approach in the animal models. |
| first_indexed | 2024-03-10T09:28:45Z |
| format | Article |
| id | doaj.art-22d495cee9bc44988de9832e90902624 |
| institution | Directory Open Access Journal |
| issn | 2076-0817 |
| language | English |
| last_indexed | 2024-03-10T09:28:45Z |
| publishDate | 2021-07-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Pathogens |
| spelling | doaj.art-22d495cee9bc44988de9832e909026242023-11-22T04:39:08ZengMDPI AGPathogens2076-08172021-07-0110789410.3390/pathogens10070894The Autofluorescence Patterns of <i>Acanthamoeba castellanii, Pseudomonas aeruginosa</i> and <i>Staphylococcus aureus</i>: Effects of Antibiotics and TetracaineHari Kumar Peguda0Saabah B. Mahbub1Tashi Doma Sherpa2Dinesh Subedi3Abbas Habibalahi4Ayad G. Anwer5Zi Gu6Mark D. P. Willcox7Ewa M. Goldys8Nicole A. Carnt9School of Optometry and Vision Science, University of New South Wales, Sydney 2052, AustraliaARC Centre of Excellence for Nanoscale Biophotonics, Graduate School of Biomedical Engineering, University of New South Wales, Sydney 2052, AustraliaSchool of Optometry and Vision Science, University of New South Wales, Sydney 2052, AustraliaSchool of Optometry and Vision Science, University of New South Wales, Sydney 2052, AustraliaARC Centre of Excellence for Nanoscale Biophotonics, Graduate School of Biomedical Engineering, University of New South Wales, Sydney 2052, AustraliaARC Centre of Excellence for Nanoscale Biophotonics, Graduate School of Biomedical Engineering, University of New South Wales, Sydney 2052, AustraliaSchool of Chemical Engineering, University of New South Wales, Sydney 2052, AustraliaSchool of Optometry and Vision Science, University of New South Wales, Sydney 2052, AustraliaARC Centre of Excellence for Nanoscale Biophotonics, Graduate School of Biomedical Engineering, University of New South Wales, Sydney 2052, AustraliaSchool of Optometry and Vision Science, University of New South Wales, Sydney 2052, Australia<i>Acanthamoeba</i> Keratitis (AK) can lead to substantial vision loss and morbidity among contact lens wearers. Misdiagnosis or delayed diagnosis is a major factor contributing to poor outcomes of AK. This study aimed to assess the effect of two antibiotics and one anaesthetic drug used in the diagnosis and nonspecific management of keratitis on the autofluorescence patterns of <i>Acanthamoeba</i> and two common bacteria that may also cause keratitis. <i>Acanthamoeba castellanii</i> ATCC 30868, <i>Pseudomonas aeruginosa</i> ATCC 9027, and <i>Staphylococcus aureus</i> ATCC 6538 were grown then diluted in either PBS (bacteria) or ¼ strength Ringer’s solution (<i>Acanthamoeba</i>) to give final concentrations of 0.1 OD at 660 nm or 10<sup>4</sup> cells/mL. Cells were then treated with ciprofloxacin, tetracycline, tetracaine, or no treatment (naïve). Excitation–emission matrices (EEMs) were collected for each sample with excitation at 270–500 nm with increments in 5 nm steps and emission at 280–700 nm at 2 nm steps using a Fluoromax-4 spectrometer. The data were analysed using MATLAB software to produce smoothed color-coded images of the samples tested. <i>Acanthamoeba</i> exhibited a distinctive fluorescence pattern compared to bacteria. The addition of antibiotics and anaesthetic had variable effects on autofluorescence. Tetracaine altered the fluorescence of all three microorganisms, whereas tetracycline did not show any effect on the fluorescence. Ciprofloxacin produced changes to the fluorescence pattern for the bacteria, but not <i>Acanthamoeba</i>. Fluorescence spectroscopy was able to differentiate <i>Acanthamoeba</i> from <i>P. aeruginosa and S. aureus</i> in vitro. There is a need for further assessment of the fluorescence pattern for different strains of <i>Acanthamoeba</i> and bacteria. Additionally, analysis of the effects of anti-amoebic drugs on the fluorescence pattern of <i>Acanthamoeba</i> and bacteria would be prudent before in vivo testing of the fluorescence diagnostic approach in the animal models.https://www.mdpi.com/2076-0817/10/7/894<i>Acanthamoeba</i>corneabacteriaautofluorescencekeratitis |
| spellingShingle | Hari Kumar Peguda Saabah B. Mahbub Tashi Doma Sherpa Dinesh Subedi Abbas Habibalahi Ayad G. Anwer Zi Gu Mark D. P. Willcox Ewa M. Goldys Nicole A. Carnt The Autofluorescence Patterns of <i>Acanthamoeba castellanii, Pseudomonas aeruginosa</i> and <i>Staphylococcus aureus</i>: Effects of Antibiotics and Tetracaine Pathogens <i>Acanthamoeba</i> cornea bacteria autofluorescence keratitis |
| title | The Autofluorescence Patterns of <i>Acanthamoeba castellanii, Pseudomonas aeruginosa</i> and <i>Staphylococcus aureus</i>: Effects of Antibiotics and Tetracaine |
| title_full | The Autofluorescence Patterns of <i>Acanthamoeba castellanii, Pseudomonas aeruginosa</i> and <i>Staphylococcus aureus</i>: Effects of Antibiotics and Tetracaine |
| title_fullStr | The Autofluorescence Patterns of <i>Acanthamoeba castellanii, Pseudomonas aeruginosa</i> and <i>Staphylococcus aureus</i>: Effects of Antibiotics and Tetracaine |
| title_full_unstemmed | The Autofluorescence Patterns of <i>Acanthamoeba castellanii, Pseudomonas aeruginosa</i> and <i>Staphylococcus aureus</i>: Effects of Antibiotics and Tetracaine |
| title_short | The Autofluorescence Patterns of <i>Acanthamoeba castellanii, Pseudomonas aeruginosa</i> and <i>Staphylococcus aureus</i>: Effects of Antibiotics and Tetracaine |
| title_sort | autofluorescence patterns of i acanthamoeba castellanii pseudomonas aeruginosa i and i staphylococcus aureus i effects of antibiotics and tetracaine |
| topic | <i>Acanthamoeba</i> cornea bacteria autofluorescence keratitis |
| url | https://www.mdpi.com/2076-0817/10/7/894 |
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