Antibacterial mechanism of Pseudomonas aeruginosa UKMP14T rhamnolipids against multidrug resistant Acinetobacter baumannii.

Rhamnolipids, a major category of glycolipid biosurfactant, have recently gained enormous attention in medical field because of their relevance as effective antibacterial agents against a wide variety of pathogenic bacteria. Our previous studies have shown that rhamnolipids from an environmental isolate of Pseudomonas aeruginosa UKMP14T possess antibacterial, anti-adhesive and anti-biofilm activity against multidrug-resistant ESKAPE (Enterococcus faecium, Staphylococcus aureus, Klebsiella pneumoniae, Acinetobacter baumannii, Pseudomonas aeruginosa and Enterobacter sp.) pathogens. However, the mechanism of their antibacterial action remains unclear. Thus, this study aimed to elucidate the mechanism of the antibacterial action of P. aeruginosa UKMP14T rhamnolipids by studying the changes in cells of one of the ESKAPE pathogens, Acinetobacter baumannii, which is the most difficult strain to kill. Results revealed that rhamnolipid treatment rendered A. baumannii cells more hydrophobic as evaluated through contact angle measurements. It also induced the release of cellular proteins measuring 510 µg/mL at a rhamnolipid concentration of 1000 µg/mL. In addition, rhamnolipids were found to be bactericidal in their action as they could permeate the inner membranes, leading to a leak-out of nucleotides. More than 50 % of the cells were found to be killed upon 1000 µg/mL rhamnolipid treatment as observed through fluorescence microscopy. Other cellular changes such as irregular shape and size, membrane perturbations, clumping, shrinkage and physical damage were clearly visible in SEM, FESEM and laser micrographs. Furthermore, rhamnolipid treatment inhibited the levels of acyl-homoserine lactones (AHLs) in A. baumannii, which are vital for their biofilm formation and virulence. The obtained results indicate that P. aeruginosa UKMP14T rhamnolipids target outer and inner bacterial membranes through permeation, including physical damage to the cells, leading to cell leakage. Furthermore, AHL inhibition appears to be the mechanism behind their anti-biofilm action. All these observations can be correlated to rhamnolipids' antibacterial effect against A. baumannii.

Antimicrobial, antiadhesive, and antibiofilm actions of rhamnolipids on ESKAPE pathogens.

The aim of this study was to test the antimicrobial, antiadhesive, and antibiofilm activities of a rhamnolipid extracted from Pseudomonas aeruginosa UKMP14T previously isolated from oil-contaminated soil in Malaysia against ESKAPE (i.e. multidrug resistant) pathogens. Zones of inhibition in an agar well diffusion assay were observed at 50 µg mL-1 concentrations of rhamnolipid for all the ESKAPE bacteria. The MIC and MBC values ranged between 7.81-62.5 µg mL-1 and 31.25-1000 µg mL-1, respectively. Percent killing was recorded to be >90% except for Klebsiella pneumoniae (86.84%). Furthermore, antiadhesion studies showed that there was 76% hindrance in attachment of Enterococcus faecium and 91% in Acinetobacter baumannii at 4 × MIC. The highest inhibition in adhesion was found at 4 × MIC, which was 46% for Ac. baumannii and 62% for Enterococcus faecium. Finally, the antibiofilm capability of the rhamnolipid was determined, which ranged between 25%-76% in Ac. baumannii and 35%-88% in Enterococcus faecium. To the best of our knowledge, this is the first study to include research on antimicrobial, antiadhesive and antibiofilm activities of rhamnolipid from the local isolate Ps. aeruginosa UKMP14T against ESKAPE bacteria. Obtained results suggest that this rhamnolipid can be exploited commercially for the production of novel antibiotics.

Optimisation and production of alpha amylase from thermophilic Bacillus spp. and its application in food waste biodegradation.

This study employed Bacillus spp. with α-amylase production isolated from Malaysian hot spring for domestic kitchen food waste treatment contained grains, vegetables, chicken and tuna that mimic the food waste discharge from domestic kitchens in Malaysian household. Results showed that Bacillus licheniformis HULUB1 and Bacillus subtilis SUNGB2 possess excellent amylolytic properties. Highest α-amylase activity was obtained when both isolates were cultivated at pH 6.0 and 65 °C with concentrations of 18.15 U/mL for HULUB1 and 22.14 U/mL for SUNGB2. Stability of α-amylase with significant levels of enzyme activity were recorded at 55-85 °C and pH 5.0-9.0. The extracted mixed α-amylase of HULUB1 and SUNGB2 showed greatest reduction were achieved at day 12 with 45% ± 0.03 solid content at 65 °C. While the mixed culture of HULUB1 and SUNGB2 displayed an enhanced effect on the food waste contents reduction with 43% ± 0.02 solid content at 45 °C after day 12. The findings showed that the combination of the two Bacillus spp. isolates possessed degradation of food wastes at faster rate than α-amylase. It was also pointed out that the standard food waste (SFW) and the treatment process assimilated for this study was suitable for the growth of Bacillus spp.

Transcriptomic Response in Pseudomonas aeruginosa towards Treatment with a Kaempferol Isolated from Melastoma malabathricum Linn Leaves.

Pseudomonas aeruginosa is one of the main causes of nosocomial infections and is frequently associated with opportunistic infections among hospitalized patients. Kaempferol-3-O-(2',6'-di-O-trans-p-coumaroyl)-ß-D glucopyranoside (K F) is an antipseudomonal compound isolated from the leaves of the native medicinal plant Melastoma malabathricum. Herein, an RNA-seq transcriptomic approach was employed to study the effect of K F treatment on P. aeruginosa and to elucidate the molecular mechanisms underlying the response to K F at two time points (6 h and 24 h incubation). Quantitative real-time PCR (qRT-PCR) was performed for four genes (uvrD, sodM, fumC1, and rpsL) to assess the reliability of the RNA-seq results. The RNA-seq transcriptomic analysis revealed that K F increases the expression of genes involved in the electron transport chain (NADH-I), resulting in the induction of ATP synthesis. Furthermore, K F also increased the expression of genes associated with ATP-binding cassette transporters, flagella, type III secretion system proteins, and DNA replication and repair, which may further influence nutrient uptake, motility, and growth. The results also revealed that K F decreased the expression of a broad range of virulence factors associated with LPS biosynthesis, iron homeostasis, cytotoxic pigment pyocyanin production, and motility and adhesion that are representative of an acute P. aeruginosa infection profile. In addition, P. aeruginosa pathways for amino acid synthesis and membrane lipid composition were modified to adapt to K F treatment. Overall, the present research provides a detailed view of P. aeruginosa adaptation and behaviour in response to K F and highlights the possible therapeutic approach of using plants to combat P. aeruginosa infections.

Mechanical characteristics of antibacterial epoxy resin adhesive wood biocomposites against skin disease.

Moldy wood can cause some skin disease. However epoxy resin adhesive (EP) can inhibit mold growth. Therefore, antibacterial EP/wood biocomposites were reinforced and analyzed by the nonlinear finite element. Results show that glass fiber cloth and aluminum foil have the obvious reinforced effect under flat pressure, but this was not the case under side pressure. And when the assemble pattern was presented in 5A way, the strengthening effect was better. The nonlinear finite element showed that the aluminum foil and glass fiber cloth have the obvious reinforced effect. The mutual influence and effect of span, thickness and length on the ultimate bearing capacity of specimen were studied. And the simulation results agreed with the test. It provided a theoretical basis on the preparation of antibacterial EP/wood biocomposites against skin disease.