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Chlorhexidine promotes psl in Pseudomonas aeruginosa that enhances cell aggregation with preserved pathogenicity demonstrates an adaptation against antiseptic

ABSTRACT: Because Pseudomonas aeruginosa is a common pathogen that frequently contacts with Chlorhexidine digluconate (a regular antiseptic), then adaptations against Chlorhexidine were tested. In com-parison with the parent strain, the Chlorhexidine-adapted strain formed smaller colonies with the downregulation of several metabolic pathways (proteomic analysis) and increased resistance against colistin (an antibiotic for the current antibiotic-resistant bacteria), partly through the modification of L-Ara4N in the lipopolysaccharide at the outer membrane (proteomic analysis). Chlorhexidine-adapted strain formed dense liquid-solid interface biofilm with cell aggregation partly due to the Chlorhexidine-induced overexpression of psl (exopolysaccharide-encoded gene) through LadS/GacSA pathway (c-di-GMP-independence) in 12 h biofilms and maintained the ag-gregation with SiaD-mediated c-di-GMP dependence in 24 h biofilms as evaluated by polymerase chain reaction (PCR). The addition of Ca2+ in the Chlorhexidine-adapted strain facilitated several Psl-associated genes, indicating an impact of Ca2+ in Psl production. The sessile Chlorhexi-dine-treated bacteria demonstrated a lower expression of IL-6 and IL-8 on fibroblasts and mac-rophages than the parent strain, indicating less inflammatory reactions from the Chlorhexidine use. However, the Chlorhexidine-treated bacteria induces similar severity in 14 days of mouse wounds as indicated by wound score and bacterial burdens. In conclusion, Chlorhexidine induced psl over-expression and colistin cross-resistance that might be clinically important.

INSTRUMENT(S): Q Exactive HF

ORGANISM(S): Homo Sapiens (human)

SUBMITTER: Jiradej Makjaroen

LAB HEAD: Asada Leelahavanichkul

PROVIDER: PXD034530 | Pride | 2022-10-14

REPOSITORIES: Pride

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Chlorhexidine Promotes Psl Expression in Pseudomonas aeruginosa That Enhances Cell Aggregation with Preserved Pathogenicity Demonstrates an Adaptation against Antiseptic.

Singkham-In Uthaibhorn U Phuengmaung Pornpimol P Makjaroen Jiradej J Saisorn Wilasinee W Bhunyakarnjanarat Thansita T Chatsuwan Tanittha T Chirathaworn Chintana C Chancharoenthana Wiwat W Leelahavanichkul Asada A

International journal of molecular sciences 20220727 15

Because Pseudomonas aeruginosa is frequently in contact with Chlorhexidine (a regular antiseptic), bacterial adaptations are possible. In comparison with the parent strain, the Chlorhexidine-adapted strain formed smaller colonies with metabolic downregulation (proteomic analysis) with the cross-resistance against colistin (an antibiotic for several antibiotic-resistant bacteria), partly through the modification of L-Ara4N in the lipopolysaccharide at the outer membrane. Chlorhexidine-adap ...[more]

PMID: 35955437

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Project description:Transcriptional profiles of Escherichia coli MG1655 in mixed culture with Pseudomonas aeruginosa PAO1 showed a number of E. coli genes to be upregulated including purA-F and other genes associated with purine synthesis. In contrast, genes associated with pyrimidine synthesis were unaffected. Competition experiments in both planktonic and biofilm cultures, using three purine synthesis mutants, purD, purH, and purT showed little difference in E. coli survival from the parent strain. As purines are components of the cell signals, cAMP and c-di-GMP, we conducted competition experiments with E. coli mutants lacking adenylate cyclase (cyaA), cAMP phosphodiesterase (cpdA), and the catabolite receptor protein (crp), as well as ydeH, an uncharacterized gene that has been associated with c-di-GMP synthesis. Survival of the cyaA and crp mutants during co-culture were significantly less than the parent strain. Supplementation of the media with 1mM cAMP could restore survival of the cyaA mutant but not the crp mutant. In contrast, survival of the cpdA mutant was similar to the parent strain. Survival of the ydeH mutant was moderately less than the parent, suggesting that cAMP has more impact on E. coli mixed culture growth than c-di-GMP. Addition of 1 mM indole restored the survival of both the cyaA and crp mutations. Mutants in genes for tryptophan synthesis (trpE) and indole production (tnaA) showed a loss of competition and recovery through indole supplementation, comparable to the cyaA and crp mutants. Overall, these results suggest indole and cAMP as major contributing factors to E. coli growth in mixed culture. Two replicates of LB grown E. coli ZK126 and P. aeruginosa PAO1 were grown in pure culture at 37C to an OD of 0.3 (log phase) and then mixed 1:1 and incubated for an additional 45 min. RNA was extracted, purified, reverse transcribed to cDNA and then analyzed on E. coli and P. aeruginosa chips from Affymetrix. Expression profiles were compared to pure culture E. coli and P. aeruginosa grown on LB.

			Action	DRS
	Bio_Set1_F.msf	Msf
	Bio_Set1_F.msfView	Msf
	Bio_Set1_F.pdResult	Other
	Bio_Set1_F.pdResultView	Other
	Bio_Set1_F1.mgf	Mgf

Dataset Information

Chlorhexidine promotes psl in Pseudomonas aeruginosa that enhances cell aggregation with preserved pathogenicity demonstrates an adaptation against antiseptic

Dataset's files

Publications

Chlorhexidine Promotes <i>Psl</i> Expression in <i>Pseudomonas aeruginosa</i> That Enhances Cell Aggregation with Preserved Pathogenicity Demonstrates an Adaptation against Antiseptic.

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