Proteomics Analysis Reveals a Potential Antibiotic Cocktail Therapy Strategy for Aeromonas hydrophila Infection in Biofilm

Abstract

Antibiotic fitness and acquired resistance are the two critical factors when bacteria respond to antibiotics, and the correlations and mechanisms between these two factors remain largely unknown. In this study, a TMT-labeling-based quantitative proteomics method was used to compare the differential expression of proteins between the fitness and acquired resistance to chlortetracycline in Aeromonas hydrophila biofilm. Bioinformatics analysis showed that translation-related ribosomal proteins, such as 30s ribosome subunits, increased in both factors; fatty acid biosynthesis related proteins, such as FabB, FabD, FabG, AccA, and AccD, increased in biofilm fitness, and some pathways (including propanoate-metabolism-related protein, such as PrpB, AtoB, PflB, AcsA, PrpD, and GabT) displayed decreased abundance in acquired resistance biofilm. The varieties of selected proteins involved in fatty acid biosynthesis and propanoate metabolism were further validated by q-PCR assay or Western blotting. Furthermore, the antibiotic-resistance-function assays showed that fatty-acid biosynthesis should be a protective antibiotics-resistance mechanism and a cocktail of chlortetracycline and triclosan, a fatty-acid-biosynthesis inhibitor, exhibited more efficient antimicrobial capability than did each antibiotic individually on biofilm, specifically on chlortetracycline-sensitive biofilm. We therefore demonstrate that the up-regulation of fatty acid biosynthesis may play an important role in antibiotic resistance and suggest that a cocktail of chlortetracycline and triclosan may be a potential cocktail therapy for pathogenic infections in biofilm.

Proteomic shifts in multi-species oral biofilms caused by Anaeroglobus geminatus

“…Bacterial ribosomal subunits are targets for many antibiotics, such as tetracycline 44 , azithromycin 45 and different other aminoglycosides 46 . Therefore, another important aspect on the ribosomal proteins is their association with antibiotic resistance 47–49 . Although the contribution of ribosomal proteins to antibiotic resistances of single bacterial species is heavily discussed 39 , there is lack of such knowledge in terms of whole biofilm communities.…”

Interactome of E. piscicida and grouper liver proteins reveals strategies of bacterial infection and host immune response

“…The two assays were performed as described previously5152. Mouse antisera against the fish immunoglobulin mu heavy chain, acetyl-Coenzyme A acyltransferase, annexin max1, ribosomal protein S16, cathepsin K, hemoglobin beta chain, ribosomal protein S26, glutaryl-CoA dehydrogenase, hyperosmotic glycine-rich protein and flavin-containing monooxygenase and against E. piscicida EvpB, ETAE_1826, OmpA, ETAE_3048 ETAE_0245, ETAE_2675 and ETAE_2572 were used as the primary antibodies, and a horseradish peroxidase (HRP)-conjugated rabbit anti-mouse antibody was used as the secondary antibody.…”

Serine protease Bm-SP142 was differentially expressed in resistant and susceptible Bombyx mori strains, involving in the defence response to viral infection

“…qPCR was performed in triplicate with 10 μl SYBR Premix Ex TaqTM II (2×), 0.2 μl PCR Forward Primer (20μM), 0.2μl PCR Reverse Primer (20μM), 0.4μl ROX Reference Dye (50×), 2 μl cDNA template and 7.2 μl ddH 2 O as described previously with a modification [18]. Cycling parameters were as follows: 95°C for 30s, followed by 40 cycles of 5 s at 95°C, 31 s at 60°C, 27 s at 72°C.…”