Application of cuminaldehyde and ciprofloxacin for the effective control of biofilm assembly of Pseudomonas aeruginosa: A combinatorial study.

Pseudomonas aeruginosa is widely associated with biofilm-mediated antibiotic resistant chronic and acute infections which constitute a persistent healthcare challenges. Addressing this threat requires exploration of novel therapeutic strategies involving the combination of natural compounds and conventional antibiotics. Hence, our study has focused on two compounds; cuminaldehyde and ciprofloxacin, which were strategically combined to target the biofilm challenge of P. aeruginosa. The minimum inhibitory concentration (MIC) of cuminaldehyde and ciprofloxacin was found to be 400 µg/mL and 0.4 µg/mL, respectively. Moreover, the fractional inhibitory concentration index (FICI = 0.62) indicated an additive interaction prevailed between cuminaldehyde and ciprofloxacin. Subsequently, sub-MIC doses of cuminaldehyde (25 µg/mL) and ciprofloxacin (0.05 µg/mL) were selected for an array of antibiofilm assays which confirmed their biofilm inhibitory potential without exhibiting any antimicrobial activity. Furthermore, selected doses of the mentioned compounds could manage biofilm on catheter surface by inhibiting and disintegrating existing biofilm. Additionally, the test combination of the mentioned compounds reduced virulence factors secretion, accumulated reactive oxygen species and increased cell-membrane permeability. Thus, the combination of cuminaldehyde and ciprofloxacin demonstrates potential in combating biofilm-associated Pseudomonal threats.

Piperine, a Plant Alkaloid, Exhibits Efficient Disintegration of the Pre-existing Biofilm of Staphylococcus aureus: a Step Towards Effective Management of Biofilm Threats.

Staphylococcus aureus causes a range of chronic infections in humans by exploiting its biofilm machinery and drug-tolerance property. Although several strategies have been proposed to eradicate biofilm-linked issues, here, we have explored whether piperine, a bioactive plant alkaloid, can disintegrate an already existing Staphylococcal biofilm. Towards this direction, the cells of S. aureus were allowed to develop biofilm first followed by treatment with the test concentrations (8 and 16 µg/mL) of piperine. In this connection, several assays such as total protein recovery assay, crystal violet assay, extracellular polymeric substances (EPS) measurement assay, fluorescein diacetate hydrolysis assay, and fluorescence microscopic image analysis confirmed the biofilm-disintegrating property of piperine against S. aureus. Piperine reduced the cellular auto-aggregation by decreasing the cell surface hydrophobicity. On further investigation, we observed that piperine could down regulate the dltA gene expression that might reduce the cell surface hydrophobicity of S. aureus. It was also observed that the piperine-induced accumulation of reactive oxygen species (ROS) could enhance biofilm disintegration by decreasing the cell surface hydrophobicity of the test organism. Together, all the observations suggested that piperine could be used as a potential molecule for the effective management of the pre-existing biofilm of S. aureus.

Combinatorial application of cuminaldehyde and gentamicin shows enhanced antimicrobial and antibiofilm action on Pseudomonas aeruginosa.

The emergence of biofilm-induced drug tolerance poses a critical challenge to public healthcare management. Pseudomonas aeruginosa, a gram-negative opportunistic bacterium, is involved in various biofilm-associated infections in human hosts. Towards this direction, in the present study, a combinatorial approach has been explored as it is a demonstrably effective strategy for managing microbial infections. Thus, P. aeruginosa has been treated with cuminaldehyde (a naturally occurring phytochemical) and gentamicin (an aminoglycoside antibiotic) in connection to the effective management of the biofilm challenges. It was also observed that the test molecules could show increased antimicrobial activity against P. aeruginosa. A fractional inhibitory concentration index (FICI) of 0.65 suggested an additive interaction between cuminaldehyde and gentamicin. Besides, a series of experiments such as crystal violet assay, estimation of extracellular polymeric substance (EPS), and microscopic images indicated that an enhanced antibiofilm activity was obtained when the selected compounds were applied together on P. aeruginosa. Furthermore, the combination of the selected compounds was found to reduce the secretion of virulence factors from P. aeruginosa. Taken together, this study suggested that the combinatorial application of cuminaldehyde and gentamicin could be considered an effective approach towards the control of biofilm-linked infections caused by P. aeruginosa.

Cuminaldehyde and Tobramycin Forestall the Biofilm Threats of Staphylococcus aureus: A Combinatorial Strategy to Evade the Biofilm Challenges.

Staphylococcus aureus, an opportunistic Gram-positive pathogen, is known for causing various infections in humans, primarily by forming biofilms. The biofilm-induced antibiotic resistance has been considered a significant medical threat. Combinatorial therapy has been considered a reliable approach to combat antibiotic resistance by using multiple antimicrobial agents simultaneously, targeting bacteria through different mechanisms of action. To this end, we examined the effects of two molecules, cuminaldehyde (a natural compound) and tobramycin (an antibiotic), individually and in combination, against staphylococcal biofilm. Our experimental observations demonstrated that cuminaldehyde (20 µg/mL) in combination with tobramycin (0.05 µg/mL) exhibited efficient reduction in biofilm formation compared to their individual treatments (p < 0.01). Additionally, the combination showed an additive interaction (fractional inhibitory concentration value 0.66) against S. aureus. Further analysis revealed that the effective combination accelerated the buildup of reactive oxygen species (ROS) and increased the membrane permeability of the bacteria. Our findings also specified that the cuminaldehyde in combination with tobramycin efficiently reduced biofilm-associated pathogenicity factors of S. aureus, including fibrinogen clumping ability, hemolysis property, and staphyloxanthin production. The selected concentrations of tobramycin and cuminaldehyde demonstrated promising activity against the biofilm development of S. aureus on catheter models without exerting antimicrobial effects. In conclusion, the combination of tobramycin and cuminaldehyde presented a successful strategy for combating staphylococcal biofilm-related healthcare threats. This combinatorial approach holds the potential for controlling biofilm-associated infections caused by S. aureus.

Isolation of a soil bacterium for remediation of polyurethane and low-density polyethylene: a promising tool towards sustainable cleanup of the environment.

A soil bacterium, designated strain AKS31, was isolated on the plastic polyurethane (PUR) and based on the molecular and biochemical analysis was tentatively assigned to the genus Pseudomonas. Preliminary studies suggested that strain AKS31 had the capability of biodegrading polyurethane (PUR) and low-density polyethylene (LDPE). This observation was confirmed by the analysis of the biodegradation products. The hydrolyzed products of PUR analyzed sequentially by High-Performance Liquid Chromatography (HPLC) and Matrix Assisted Laser Desorption Ionization Time of Flight Mass Spectrometry (MALDI-TOF MS) showed the presence of diethylene glycol suggesting the presence of an esterase. A gene that could be involved in producing an esterase-like activity (PURase gene) was identified after the amplification and sequencing of a PCR product. Fourier Transformed Infrared (FTIR) spectrophotometric analysis of AKS31-treated LDPE film revealed the incorporation of hydroxyl groups suggesting the involvement of a hydroxylase in the degradation of LDPE. It is established that plastics form microplastics and microbeads in soils which negatively impact the health of living organisms and there have been concentrated research efforts to remediate this problem. Microcosm studies revealed that when strain AKS31 was bioaugmented with soil both the polymers were degraded during which time the heterotrophic plate counts, soil respiration and soil organic carbon content increased but this was not the case with the control nonbioaugmented microcosm. The results demonstrate that the strain AKS31 may have the potential in biodegradation of PUR and LPDE present as plastic microbeads and thereby improving soil health. Further studies in this direction are warranted. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1007/s13205-020-02592-9.

Nitrogen dynamics of the aquatic system is an important driving force for efficient sewage purification in single pond natural treatment wetlands at East Kolkata Wetland.

At East Kolkata Wetlands, though the domestic city sewage is purified very rapidly, the mechanisms of treatment remains inadequately explored. In this context, the present study investigated nitrogen dynamics of the single pond treatment systems during purification and explored its potential role in sewage treatment. For this purpose the concentrations of different forms of nitrogen present both in water and soil at different time points of purification were measured. The organic nitrogen content decreased sharply, in the early phase, with an increase in ammonium concentration. Notably the reduction in organic nitrogen was significantly higher than the increase in NH4+ which can be attributed to the volatilization of NH4+ under alkaline pH. This volatilization results in reduced oxygen demand. The nitrate-N concentration decreased sharply from soil with a concomitant increase in water column. However the reduction of nitrate in soil was significantly higher than the increase in water column. It indicated the occurrence of denitrification under anoxic condition wherein nitrate serves as terminal electron acceptor. Additionally a part of the nitrate supported planktonic growth. Thus it describes another mechanism of reducing oxygen demand. The initial NH4+-N concentration in the soil was very low and it increased gradually during purification due to increasing soil cation exchange capacity. Thus by trapping NH4+ ion soil contributes towards preventing contamination of water. Thus at EKW, the cumulative activities in water and soil involved in nitrogen dynamics lead to overall reduction of the oxygen demand and contribute towards efficient sewage purification.