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 phytochemical prevents the biofilm city of methicillin-resistant Staphylococcus aureus: A biochemical approach to understand the underlying mechanism.

Methicillin-resistant Staphylococcus aureus (MRSA), a drug-resistant human pathogen causes several nosocomial as well as community-acquired infections involving biofilm machinery. Hence, it has gained a wide interest within the scientific community to impede biofilm-induced MRSA-associated health complications. The current study focuses on the utilization of a natural bioactive compound called piperine to control the biofilm development of MRSA. Quantitative assessments like crystal violet, total protein recovery, and fluorescein-di-acetate (FDA) hydrolysis assays, demonstrated that piperine (8 and 16 µg/mL) could effectively compromise the biofilm formation of MRSA. Light and scanning electron microscopic image analysis confirmed the same. Further investigation revealed that piperine could reduce extracellular polysaccharide production by down-regulating the expression of icaA gene. Besides, piperine could reduce the cell-surface hydrophobicity of MRSA, a crucial factor of biofilm formation. Moreover, the introduction of piperine could interfere with microbial motility indicating the interaction of piperine with the quorum-sensing components. A molecular dynamics study showed a stable binding between piperine and AgrA protein (regulator of quorum sensing) suggesting the possible meddling of piperine in quorum-sensing of MRSA. Additionally, the exposure to piperine led to the accumulation of intracellular reactive oxygen species (ROS) and potentially heightened cell membrane permeability in inhibiting microbial biofilm formation. Besides, piperine could reduce the secretion of diverse virulence factors from MRSA. Further exploration revealed that piperine interacted with extracellular DNA (e-DNA), causing disintegration by weakening the biofilm architecture. Conclusively, this study suggests that piperine could be a potential antibiofilm molecule against MRSA-associated biofilm infections.

Bioaugmentation of Enterobacter cloacae AKS7 causes an enhanced degradation of low-density polyethylene (LDPE) in soil: a promising approach for the sustainable management of LDPE waste.

Enterobacter cloacae AKS7 was previously reported to degrade UV-treated low-density polyethylene (LDPE) more efficiently than UV-untreated LDPE. However, the degradation of LDPE by Enterobacter cloacae AKS7 at the LDPE-contaminated soil remained unaddressed. To address this issue, soil microcosms were prepared in which an equal amount of either UV-treated or UV-untreated LDPE was added. Then, the microcosms were either augmented with AKS7 or left non-augmented. We observed that the bioaugmented microcosms exhibited approximately twofold greater polymer degradation than non-bioaugmented microcosms. To investigate the underlying cause, we found that the abundance of LDPE-degrading organisms got increased by approximately fivefold in bioaugmented microcosms than non-bioaugmented microcosms. The microbial biomass carbon and nitrogen content got enhanced by approximately twofold in bioaugmented microcosms as contrasted to non-bioaugmented microcosms. Furthermore, the bioaugmented microcosms showed almost twofold increase in the level of dehydrogenase and fluorescein diacetate (FDA) hydrolyzing activity than the non-bioaugmented microcosms. To add on, Shannon-diversity index and Gini coefficient were determined in each microcosm to measure the microbial richness and evenness, respectively, using the results of carbon source utilization pattern of BiOLOG ECO plate. The bioaugmented microcosms exhibited ~ 30% higher functional richness and ~ 30% enhanced functional evenness than the non-bioaugmented microcosms indicating the formation of an enriched ecosystem that could offer various functions including polymer degradation. Taken together, the results suggested that Enterobacter cloacae AKS7 could be used as a promising bioaugmenting agent for the sustainable degradation of LDPE waste at a contaminated site.

Degradation of low-density poly ethylene (LDPE) by Enterobacter cloacae AKS7: a potential step towards sustainable environmental remediation.

Plastics composed of polyethylene are non-biodegradable and are mostly harmful to the environment. Literature studies documented that the extent of microbial degradation of low-density polyethylene (LDPE) seems to be insufficient and the underlying mechanisms of such degradation remain unexplored. In the present study, efforts were given to degrade LDPE by a recently isolated bacteria Enterobacter cloacae AKS7. Scanning electron microscopic (SEM) image, tensile strength, and weight loss analysis confirmed the efficient degradation of LDPE by AKS7. To investigate the mechanism, it was observed that with the progression of time, the extent of microbial colonization got increased considerably over the LDPE surface. It was also observed that the organism (AKS7) gradually increased the secretion of extracellular polymeric substances (EPS) suggesting the formation of efficient biofilm over the LDPE surface. Furthermore, to comprehend the role of cell-surface hydrophobicity towards biofilm formation, two mutants of AKS7 were screened that showed a considerable reduction in cell-surface hydrophobicity in contrast to its wild type. The result showed that the mutants revealed compromised LDPE degradation than wild-type cells of AKS7. Further investigation revealed that the mutant cells of AKS7 were incapable of adhering to LDPE in contrast to wild-type cells. Thus, the results demonstrated that the cell-surface hydrophobicity of AKS7 favors the development of microbial biofilm over LDPE that leads to the enhanced degradation of LDPE by AKS7. Therefore, the organism holds the assurance to be considered as a promising bio-remediating agent for the sustainable degradation of polythene-based hazardous waste.

Inhibition of biofilm formation of Pseudomonas aeruginosa by caffeine: a potential approach for sustainable management of biofilm.

Pseudomonas aeruginosa is a potent biofilm forming organism causing several diseases on host involving biofilm. Several natural and synthetic molecules have been explored towards inhibiting the biofilm formation of Pseudomonas aeruginosa. In the current report, the role of a natural molecule namely caffeine was examined against the biofilm forming ability of P. aeruginosa. We have observed that caffeine shows substantial antimicrobial activity against P. aeruginosa wherein the minimum inhibitory concentration (MIC) of caffeine was found to be 200 µg/mL. The antibiofilm activity of caffeine was determined by performing a series of experiments using its sub-MIC concentrations (40 and 80 µg/mL). The results revealed that caffeine can significantly inhibit the biofilm development of P. aeruginosa. Caffeine has been found to interfere with the quorum sensing of P. aeruginosa by targeting the swarming motility. Molecular docking analysis further indicated that caffeine can interact with the quorum sensing proteins namely LasR and LasI. Thus, the result indicated that caffeine could inhibit the formation of biofilm by interfering with the quorum sensing of the organism. Apart from biofilm inhibition, caffeine has also been found to reduce the secretion of virulence factors from Pseudomonas aeruginosa. Taken together, the results revealed that in addition to biofilm inhibition, caffeine can also decrease the spreading of virulence factors from Pseudomonas 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.

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 exhibits potential antibiofilm activity against Pseudomonas aeruginosa involving reactive oxygen species (ROS) accumulation: a way forward towards sustainable biofilm management.

Pseudomonas aeruginosa often causes various acute and chronic infections in humans exploiting biofilm. Molecules interfering with microbial biofilm formation could be explored for the sustainable management of infections linked to biofilm. Towards this direction, the antimicrobial and antibiofilm activity of cuminaldehyde, an active ingredient of the essential oil of Cuminum cyminum was tested against Pseudomonas aeruginosa. In this regard, the minimum inhibitory concentration (MIC) of cuminaldehyde was found to be 150 µg/mL against the test organism. Experiments such as crystal violet assay, estimation of total biofilm protein, fluorescence microscopy and measurement of extracellular polymeric substances (EPS) indicated that the sub-MIC doses (up to 60 µg/mL) of cuminaldehyde demonstrated considerable antibiofilm activity without showing any antimicrobial activity to the test organism. Moreover, cuminaldehyde treatment resulted in substantial accumulation of cellular reactive oxygen species (ROS) that led to the inhibition of microbial biofilm formation. To this end, the exposure of ascorbic acid was found to restore the biofilm-forming ability of the cuminaldehyde-treated cells. Besides, a noticeable reduction in proteolytic activity was also observed when the organism was treated with cuminaldehyde. Taken together, the results demonstrated that cuminaldehyde could be used as a promising molecule to inhibit the biofilm formation of Pseudomonas aeruginosa.

Bioaugmentation of soil with Enterobacter cloacae AKS7 enhances soil nitrogen content and boosts soil microbial functional-diversity.

Biofertilizer happens to be a promising alternative of chemical fertilizer in the establishment of sustainable agricultural practices. Following this observation, several nitrogen-fixing bacteria were isolated from the soil in which an isolate (AKS7) was selected for further studies as AKS7 showed considerable competence in growth on nitrogen-free growth medium. Acetylene reduction assay confirmed that AKS7 can fix atmospheric nitrogen efficiently. The result of Kjeldahl assay revealed that the organism (AKS7) could fix nitrogen up to 12 mg in 8 days. A strong positive correlation (r = 0.987) was observed between microbial cell biomass and the amount of nitrogen fixed by AKS7 over a period of 8 days. The organism was identified as Enterobacter cloacae through molecular and biochemical tests. To investigate the in situ nitrogen fixation by E. cloacae AKS7, naturally attenuated (AKS7 not-inoculated) and bioaugmented (AKS7-inoculated) soil microcosms were prepared. The bioaugmented microcosm showed higher level of soil nitrogen content than naturally attenuated microcosm. A large number of heterotrophic as well as nitrogen-fixing microorganisms were counted in bioaugmented microcosm than naturally attenuated microcosm. Results of the carbon source utilization patterns of BiOLOG ECO plate revealed that bioaugmented microcosm exhibited higher level of functional richness and evenness that lead to the exhibition of higher level of microbial functional-diversity in bioaugmented microcosm than the naturally attenuated microcosm. Taken together, the results indicated that augmentation of E. cloacae AKS7 into soil enhanced the nitrogen content and soil microbial functional-diversity considerably.