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Volume 13
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10.3390/app132111918
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Article

Virulence-Linked Mutations in Rubredoxin Reductase and Glutaredoxin: Impact on Antibiotic Susceptibility and Phage Therapy in Pseudomonas aeruginosa

by
Sara Sá
1,2,3,4,†,
Carina Silva
1,2,5,†,
Maria Clara Dias
6,
Marlene Veiga
6,
Sofia Lopes
6,
Ruben Fernandes
1,2,3,
Ana Catarina Rocha
1,2,4,
Patrick J. Pais
1,2,7,
Marco Oliveira
1,2,6,
João Mendes
1,2,
Gonçalo Novais
1,2,
Carla Luís
1,2,4,
Álvaro Gestoso
1,2,
José Mário Macedo
1,2,5,
Daniela Martins-Mendes
1,2,3,
Ana Cláudia Pereira
1,2,3,* and
Pilar Baylina
1,2,6,*
1
FP-BHS—Biomedical and Health Sciences Research Unit, FP-I3ID—Instituto de Investigação, Inovação e Desenvolvimento, FFP—Fundação Fernando Pessoa, 4249-004 Porto, Portugal
2
CECLIN—Center of Clinical Studies, HE-FP—Hospital Fernando Pessoa, 4420-096 Gondomar, Portugal
3
FCS—Faculty of Health Sciences, Fernando Pessoa University, 4200-253 Porto, Portugal
4
FMUP—Faculty of Medicine of the University of Porto, 4200-319 Porto, Portugal
5
Faculty of Biology, Department of Functional Biology and Health Sciences, University of Vigo, 36310 Vigo, Spain
6
ESS—School of Health, IPP—Polytechnic Institute of Porto, 4200-072 Porto, Portugal
7
ECVA—School of Life and Environmental Sciences, UTAD—University of Trás-os-Montes and Alto Douro, 5000-801 Vila Real, Portugal
*
Authors to whom correspondence should be addressed.
†
These authors contributed equally to this work.
Appl. Sci. 2023, 13(21), 11918; https://doi.org/10.3390/app132111918
Submission received: 1 September 2023 / Revised: 29 September 2023 / Accepted: 10 October 2023 / Published: 31 October 2023
(This article belongs to the Section Applied Biosciences and Bioengineering)
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Abstract

Pseudomonas aeruginosa (PAO1) is an opportunistic pathogen, lethal in immunocompromised individuals. The clinical management of PAO1 infections still depends deeply on antibiotic therapy. However, this therapy has been alarmingly overpowered by growing bacterial resistance mechanisms over the years. One of these bacterial mechanisms is quorum sensing (QS). QS is involved in the production of biofilm, rhamnolipids and pyocyanin, among other factors. The present study aimed to study the effect of the mutations in the genes of rubredoxin (Rub A1 and Rub A2) and glutaredoxin (GLRx) in the production of virulence traits and susceptibility of PAO1 to the antibiotic ciprofloxacin (CIP) and to infection by a phage cocktail. Rub A1, Rub A2, and GLRx showed a decrease in the expression of genes lasI, lasR, mvfR, and rpsL when compared to the wild type, PAO1. Rub A1 and Rub A2 also showed a decrease in the expression of the gene pqsA, while the mutant GLRx showed an increase of over 200% in expression compared to PAO1. The biofilm produced by the mutants Rub A1, Rub A2, and GLRx increased more than 1.5 times in comparison to PAO1, with statistical significance (p < 0.0001). In the viability assay, the mutant strain Rub A2 was the most susceptible to ciprofloxacin in both concentrations tested (p < 0.0001). The production of proteases increased in the mutant strains when compared to PAO1 (p < 0.05). However, there was a decrease in the production of rhamnolipids and pyocyanins in the mutant strains. In the phage assay, we could perceive a reduction in the growth of the mutant strains when compared to PAO1. Additionally, after the addition of the phages, all the strains showed susceptibility to the phage assay (p < 0.0001), observed in the decrease in the absorbance values. These results may highlight the relevance of the genes Rub A1, Rub A2, and GLRX in the proliferation and treatment of infections with PAO1. Overall, this study gives preliminary insights into how gene expression may be helpful in strategies to overcome antibiotic resistance.
Keywords: quorum sensing; oxidative stress; antibiotic stress; biofilm production; virulence quorum sensing; oxidative stress; antibiotic stress; biofilm production; virulence

Share and Cite

MDPI and ACS Style

Sá, S.; Silva, C.; Dias, M.C.; Veiga, M.; Lopes, S.; Fernandes, R.; Rocha, A.C.; Pais, P.J.; Oliveira, M.; Mendes, J.; et al. Virulence-Linked Mutations in Rubredoxin Reductase and Glutaredoxin: Impact on Antibiotic Susceptibility and Phage Therapy in Pseudomonas aeruginosa. Appl. Sci. 2023, 13, 11918. https://doi.org/10.3390/app132111918

AMA Style

Sá S, Silva C, Dias MC, Veiga M, Lopes S, Fernandes R, Rocha AC, Pais PJ, Oliveira M, Mendes J, et al. Virulence-Linked Mutations in Rubredoxin Reductase and Glutaredoxin: Impact on Antibiotic Susceptibility and Phage Therapy in Pseudomonas aeruginosa. Applied Sciences. 2023; 13(21):11918. https://doi.org/10.3390/app132111918

Chicago/Turabian Style

Sá, Sara, Carina Silva, Maria Clara Dias, Marlene Veiga, Sofia Lopes, Ruben Fernandes, Ana Catarina Rocha, Patrick J. Pais, Marco Oliveira, João Mendes, and et al. 2023. "Virulence-Linked Mutations in Rubredoxin Reductase and Glutaredoxin: Impact on Antibiotic Susceptibility and Phage Therapy in Pseudomonas aeruginosa" Applied Sciences 13, no. 21: 11918. https://doi.org/10.3390/app132111918

APA Style

Sá, S., Silva, C., Dias, M. C., Veiga, M., Lopes, S., Fernandes, R., Rocha, A. C., Pais, P. J., Oliveira, M., Mendes, J., Novais, G., Luís, C., Gestoso, Á., Macedo, J. M., Martins-Mendes, D., Pereira, A. C., & Baylina, P. (2023). Virulence-Linked Mutations in Rubredoxin Reductase and Glutaredoxin: Impact on Antibiotic Susceptibility and Phage Therapy in Pseudomonas aeruginosa. Applied Sciences, 13(21), 11918. https://doi.org/10.3390/app132111918

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