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1.
Mol Cell ; 81(4): 756-766.e8, 2021 02 18.
Artigo em Inglês | MEDLINE | ID: mdl-33472056

RESUMO

Bacillus subtilis structural maintenance of chromosomes (SMC) complexes are topologically loaded at centromeric sites adjacent to the replication origin by the partitioning protein ParB. These ring-shaped ATPases then translocate down the left and right chromosome arms while tethering them together. Here, we show that the site-specific recombinase XerD, which resolves chromosome dimers, is required to unload SMC tethers when they reach the terminus. We identify XerD-specific binding sites in the terminus region and show that they dictate the site of unloading in a manner that depends on XerD but not its catalytic residue, its partner protein XerC, or the recombination site dif. Finally, we provide evidence that ParB and XerD homologs perform similar functions in Staphylococcus aureus. Thus, two broadly conserved factors that act at the origin and terminus have second functions in loading and unloading SMC complexes that travel between them.


Assuntos
Bacillus subtilis/enzimologia , Proteínas de Bactérias/metabolismo , Cromossomos Bacterianos/metabolismo , Integrases/metabolismo , Staphylococcus aureus/enzimologia , Bacillus subtilis/genética , Proteínas de Bactérias/genética , Cromossomos Bacterianos/genética , DNA Primase/genética , DNA Primase/metabolismo , Integrases/genética , Staphylococcus aureus/genética
2.
Microb Pathog ; 190: 106624, 2024 May.
Artigo em Inglês | MEDLINE | ID: mdl-38492828

RESUMO

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.


Assuntos
Antibacterianos , Benzaldeídos , Biofilmes , Ciprofloxacina , Testes de Sensibilidade Microbiana , Pseudomonas aeruginosa , Espécies Reativas de Oxigênio , Biofilmes/efeitos dos fármacos , Ciprofloxacina/farmacologia , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/fisiologia , Antibacterianos/farmacologia , Benzaldeídos/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Fatores de Virulência , Cimenos/farmacologia , Sinergismo Farmacológico , Permeabilidade da Membrana Celular/efeitos dos fármacos , Humanos
3.
Microb Pathog ; 189: 106601, 2024 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-38423404

RESUMO

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.


Assuntos
Alcaloides , Benzodioxóis , Staphylococcus aureus Resistente à Meticilina , Piperidinas , Alcamidas Poli-Insaturadas , Infecções Estafilocócicas , Humanos , Antibacterianos/farmacologia , Antibacterianos/metabolismo , Biofilmes , Compostos Fitoquímicos/farmacologia , DNA/metabolismo , Testes de Sensibilidade Microbiana
4.
J Appl Microbiol ; 133(5): 3059-3068, 2022 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-35929359

RESUMO

AIMS: Psychrotrophs are extremophilic microorganisms that grow optimally in low temperature having many unique bioactive molecules of biotechnological applications. In this study, we characterized a pigment from an arctic bacterium with protective activity towards UV exposure. METHODS AND RESULTS: The present research reports isolation and characterization of a psychrotrophic bacteria, RSAP2, from the soil sample of NyAlesund (78°56"N, 11°54"E), Svalbard, Norway. The strain showed closest 16S rRNA gene sequence similarity (99.9%) with Kocuria indica NIO-1021. RSAP2 is a Gram-positive, coccoid aerobe which produces a yellow pigment. The optimal parameters for pigment production while grown in LB medium were 3% (w/v) NaCl and 4 days of incubation of the culture at 20°C and pH 9 with shaking (180 rpm). The pigment was extracted in methanol and acetone (2:1) and further purified through column chromatography. It was characterized by mass spectrometry, UV-visible, fluorescence, IR, 1 H NMR, 13 C NMR spectroscopy and CHNS/O analysis. The pigment has a molecular weight of about 258 daltons and the molecular formula was determined as C15 H18 N2 O2 and is a quinoline derivative. We show that the pigment can protect Escherichia coli against UV-mediated mutagenesis. We further demonstrate that the pigment displays a significant antimicrobial effect and in sublethal concentrations it impairs biofilm formation ability of the model organism Staphylococcus aureus. CONCLUSIONS: The pigment of a psychrotrophic Arctic bacterium, most likely a strain of K. indica, was purified and its chemical structure was determined. The quinoline-based pigment has the ability to protect live cells from UV induced damage. SIGNIFICANCE AND IMPACT OF STUDY: Analysis and characterization of this newly isolated quinoline-based pigment is a potential candidate for future application in skin care products.


Assuntos
Anti-Infecciosos , Quinolinas , RNA Ribossômico 16S/genética , Cloreto de Sódio , Metanol , Acetona , Bactérias/genética , Quinolinas/farmacologia , Solo , Filogenia , Análise de Sequência de DNA , DNA Bacteriano/genética , Ácidos Graxos/análise , Regiões Árticas , Técnicas de Tipagem Bacteriana
5.
Proc Natl Acad Sci U S A ; 116(41): 20489-20499, 2019 10 08.
Artigo em Inglês | MEDLINE | ID: mdl-31548377

RESUMO

To separate replicated sister chromatids during mitosis, eukaryotes and prokaryotes have structural maintenance of chromosome (SMC) condensin complexes that were recently shown to organize chromosomes by a process known as DNA loop extrusion. In rapidly dividing bacterial cells, the process of separating sister chromatids occurs concomitantly with ongoing transcription. How transcription interferes with the condensin loop-extrusion process is largely unexplored, but recent experiments have shown that sites of high transcription may directionally affect condensin loop extrusion. We quantitatively investigate different mechanisms of interaction between condensin and elongating RNA polymerases (RNAPs) and find that RNAPs are likely steric barriers that can push and interact with condensins. Supported by chromosome conformation capture and chromatin immunoprecipitation for cells after transcription inhibition and RNAP degradation, we argue that translocating condensins must bypass transcribing RNAPs within ∼1 to 2 s of an encounter at rRNA genes and within ∼10 s at protein-coding genes. Thus, while individual RNAPs have little effect on the progress of loop extrusion, long, highly transcribed operons can significantly impede the extrusion process. Our data and quantitative models further suggest that bacterial condensin loop extrusion occurs by 2 independent, uncoupled motor activities; the motors translocate on DNA in opposing directions and function together to enlarge chromosomal loops, each independently bypassing steric barriers in their path. Our study provides a quantitative link between transcription and 3D genome organization and proposes a mechanism of interactions between SMC complexes and elongating transcription machinery relevant from bacteria to higher eukaryotes.


Assuntos
Adenosina Trifosfatases/metabolismo , Bacillus subtilis/metabolismo , Proteínas de Bactérias/metabolismo , Cromossomos Bacterianos , Proteínas de Ligação a DNA/metabolismo , RNA Polimerases Dirigidas por DNA/metabolismo , Genoma Bacteriano , Complexos Multiproteicos/metabolismo , RNA Ribossômico/metabolismo , Transcrição Gênica , Adenosina Trifosfatases/química , Adenosina Trifosfatases/genética , Bacillus subtilis/genética , Proteínas de Bactérias/química , Proteínas de Bactérias/genética , Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/genética , RNA Polimerases Dirigidas por DNA/química , RNA Polimerases Dirigidas por DNA/genética , Complexos Multiproteicos/química , Complexos Multiproteicos/genética , Ligação Proteica , RNA Ribossômico/química , RNA Ribossômico/genética
6.
Arch Microbiol ; 203(3): 1183-1193, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33230594

RESUMO

Staphylococcus aureus, a Gram-positive opportunistic microorganism, promotes pathogenicity in the human host through biofilm formation. Microorganisms associated with biofilm often exhibit drug-resistance property that poses a major threat to public healthcare. Thus, the exploration of new therapeutic approaches is the need of the hour to manage biofilm-borne infections. In the present study, efforts are put together to test the antimicrobial as well as antibiofilm activity of 1,4-naphthoquinone against Staphylococcus aureus. The result showed that the minimum bactericidal concentration (MBC) of this compound was found to be 100 µg/mL against Staphylococcus aureus. In this regard, an array of experiments (crystal violet, biofilm protein measurement, and microscopic analysis) related to biofilm assay were conducted with the sub-MBC concentrations (1/20 and 1/10 MBC) of 1,4-naphthoquinone. All the results of biofilm assay demonstrated that these tested concentrations (1/20 and 1/10 MBC) of the compound (1,4-naphthoquinone) showed a significant reduction in biofilm development by Staphylococcus aureus. Moreover, the tested concentrations (1/20 and 1/10 MBC) of the compound (1,4-naphthoquinone) were able to reduce the microbial motility of Staphylococcus aureus that might affect the development of biofilm. Further studies revealed that the treatment of 1,4-naphthoquinone to the organism was found to increase the cellular accumulation of reactive oxygen species (ROS) that resulted in the inhibition of biofilm formation by Staphylococcus aureus. Hence, it can be concluded that 1,4-naphthoquinone might be considered as a promising compound towards biofilm inhibition caused by Staphylococcus aureus.


Assuntos
Biofilmes/efeitos dos fármacos , Naftoquinonas/farmacologia , Espécies Reativas de Oxigênio/metabolismo , Staphylococcus aureus/efeitos dos fármacos , Antibacterianos/farmacologia , Humanos , Testes de Sensibilidade Microbiana , Infecções Estafilocócicas/prevenção & controle
7.
Arch Microbiol ; 203(8): 4981-4992, 2021 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-34272991

RESUMO

Staphylococcus aureus causes several nosocomial and community-acquired infections in human host involving biofilm. Thus, strategies need to be explored to curb biofilm threats by either inhibiting the formation of biofilm or disintegrating the pre-existing biofilm. Towards this direction, we had already revealed the biofilm inhibiting properties of 1,4-naphthoquinone against S. aureus. In this study, we have investigated whether this compound can act on pre-existing biofilm. Hence, biofilm of S. aureus was developed first and challenged further with 1,4-naphthoquinone. Experiments such as crystal violet assay, fluorescence microscopy, and estimation of total biofilm protein were performed to confirm the biofilm disintegration properties of 1,4-naphthoquinone. The disintegration of pre-existing biofilm could be attributed to the generation of reactive oxygen species (ROS). To investigate further, we observed that extracellular DNA (eDNA) was found to play an important role in holding the biofilm network as DNaseI treatment could cause an efficient disintegration of the same. To examine the effect of ROS on the eDNA, we exposed pre-existing biofilm to either 1,4-naphthoquinone or a combination of both 1,4-naphthoquinone and ascorbic acid for different length of time. Post-incubation, ROS generation and the amount of eDNA associated with the biofilm were determined wherein an inversely proportional relationship was observed between them. The result indicated that with the increase of ROS generation, the amount of eDNA associated with biofilm got decreased substantially. Thus, the results indicated that the generation of ROS could degrade the eDNA thereby compromising the integrity of biofilm which lead to the disintegration of pre-existing biofilm.


Assuntos
Infecções Estafilocócicas , Staphylococcus aureus , Biofilmes , Humanos , Naftoquinonas , Espécies Reativas de Oxigênio , Staphylococcus aureus/genética
8.
Arch Microbiol ; 204(1): 59, 2021 Dec 23.
Artigo em Inglês | MEDLINE | ID: mdl-34940904

RESUMO

Staphylococcus aureus causes numerous community-acquired and nosocomial infections in humans by exploiting biofilm. In this context, this study aims to impede the formation of Staphylococcus aureus biofilm by exposing the cells to a plant-based alkaloid, piperine. Our study revealed that piperine exhibited considerable antimicrobial activity against the test organism. However, we had tested the lower concentrations (up to 32 µg/mL) of piperine to observe whether they could show any antibiofilm activity against the same organism. Several experiments, like crystal violet (CV) assay, estimation of total biofilm protein, and fluorescence microscopic observations, established that lower concentrations (up to 16 µg/mL) of piperine showed efficient antibiofilm activity against Staphylococcus aureus. In this connection, we also noticed that the lower concentrations (8 and 16 µg/mL) of piperine showed a considerable reduction in microbial metabolic activity. Besides, it was also observed that the mentioned concentrations of piperine did not compromise the microbial growth of the target organism while exhibiting antibiofilm activity. To understand the underlying mechanism of microbial biofilm inhibition under the influence of piperine, we observed that the compound was found to accumulate reactive oxygen species in the bacterial cells that could play an important role in the inhibition of biofilm formation. Furthermore, the tested concentrations (8 and 16 µg/mL) of piperine were able to inhibit the motility of the test organism that might compromise the development of biofilm. Thus, piperine could be considered as a potential agent for the effective management of biofilm threat caused by Staphylococcus aureus.


Assuntos
Alcaloides , Staphylococcus aureus , Alcaloides/farmacologia , Antibacterianos/farmacologia , Benzodioxóis , Biofilmes , Humanos , Testes de Sensibilidade Microbiana , Piperidinas , Alcamidas Poli-Insaturadas , Espécies Reativas de Oxigênio
9.
Arch Microbiol ; 202(8): 2117-2125, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32506149

RESUMO

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.


Assuntos
Enterobacter cloacae/genética , Enterobacter cloacae/metabolismo , Recuperação e Remediação Ambiental , Polietileno/metabolismo , Aderência Bacteriana/genética , Biodegradação Ambiental , Biofilmes , Genes Bacterianos/genética , Interações Hidrofóbicas e Hidrofílicas , Microscopia Eletrônica de Varredura , Mutação
10.
Arch Microbiol ; 202(3): 623-635, 2020 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-31773197

RESUMO

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.


Assuntos
Antibacterianos/farmacologia , Biofilmes/efeitos dos fármacos , Cafeína/farmacologia , Pseudomonas aeruginosa/fisiologia , Proteínas de Bactérias/genética , Proteínas de Bactérias/metabolismo , Biofilmes/crescimento & desenvolvimento , Testes de Sensibilidade Microbiana , Simulação de Acoplamento Molecular , Pseudomonas aeruginosa/efeitos dos fármacos , Pseudomonas aeruginosa/genética , Percepção de Quorum/efeitos dos fármacos , Fatores de Virulência/genética , Fatores de Virulência/metabolismo
11.
Environ Monit Assess ; 192(4): 245, 2020 Mar 20.
Artigo em Inglês | MEDLINE | ID: mdl-32198540

RESUMO

Several strategies were undertaken to increase the fertility of landfill soil as rapid urbanization remarkably decreases the agricultural land, posing challenges to the fast-growing human population. Towards this direction, soil microcosms were prepared wherein the addition of nutrient or biofertilizer or the combination of both increased the soil nitrogen and phosphate content considerably. The maximum amount of nitrogen fixation and phosphate solubilization occurred in microcosm treated with biofertilizer and nutrient. To investigate the underlying cause, we observed that separate application of nutrient or biofertilizer or combined application of both increased the abundance of nitrogen-fixing and phosphate-solubilizing bacteria in the microcosms. However, the highest abundance of nitrogen-fixing and phosphate-solubilizing bacteria was spotted in a microcosm challenged with nutrient and biofertilizer together. It was detected that with increasing population of nitrogen-fixing and phosphate-solubilizing bacteria, the soil nitrogen and phosphate level also got enhanced, respectively, thus establishing a strong positive correlation between them. The microcosm treated with biofertilizer and nutrient manifested the highest degree of heterotrophic microbial growth and microbial activity than the microcosms either treated with nutrient or biofertilizer. The microcosm treated with nutrient and biofertilizer was found to exhibit the highest functional diversity compared to others. A surface plot was constructed to demonstrate the association among microbial activity, functional diversity, and the availability of soil nitrogen and phosphate content of soil. The result indicates that the combined application of nutrient and biofertilizer increases the microbial activity leading to the formation of a heterogeneous ecosystem that enhances the nitrogen and phosphate content of landfill soil considerably.


Assuntos
Nitrogênio , Fosfatos , Solo , Resíduos Sólidos , Nitrogênio/análise , Nitrogênio/metabolismo , Fosfatos/análise , Fosfatos/metabolismo , Solo/química , Microbiologia do Solo , Instalações de Eliminação de Resíduos
12.
Appl Biochem Biotechnol ; 196(3): 1272-1291, 2024 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-37389724

RESUMO

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.


Assuntos
Alcaloides , Piperidinas , Alcamidas Poli-Insaturadas , Infecções Estafilocócicas , Humanos , Staphylococcus aureus , Biofilmes , Alcaloides/farmacologia , Benzodioxóis/farmacologia , Antibacterianos/farmacologia , Testes de Sensibilidade Microbiana
13.
Artigo em Inglês | MEDLINE | ID: mdl-38526664

RESUMO

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.

14.
Folia Microbiol (Praha) ; 68(1): 151-163, 2023 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-36192618

RESUMO

Pseudomonas aeruginosa, an opportunistic pathogen, has been found to cause several chronic and acute infections in human. Moreover, it often shows drug-tolerance and poses a severe threat to public healthcare through biofilm formation. In this scenario, two molecules, namely, cuminaldehyde and tobramycin, were used separately and in combination for the efficient management of biofilm challenge. The minimum inhibitory concentration (MIC) of cuminaldehyde and tobramycin was found to be 150 µg/mL and 1 µg/mL, respectively, against Pseudomonas aeruginosa. The checkerboard assay revealed that the fractional inhibitory concentration (FIC) index of cuminaldehyde and tobramycin was 0.36 suggesting a synergistic association between them. The sub-MIC dose of cuminaldehyde (60 µg/mL) or tobramycin (0.06 µg/mL) individually did not show any effect on the microbial growth curve. However, the same combinations could affect microbial growth curve of Pseudomonas aeruginosa efficiently. In connection to biofilm management, it was observed that the synergistic interaction between cuminaldehyde and tobramycin could inhibit biofilm formation more efficiently than their single use (p < 0.01). Further investigation revealed that the combinations of cuminaldehyde and tobramycin could generate reactive oxygen species (ROS) that resulted in the increase of membrane permeability of bacterial cells leading to the efficient inhibition of microbial biofilm formation. Besides, the synergistic interaction between cuminaldehyde (20 µg/mL) and tobramycin (0.03 µg/mL) also showed significant biofilm dispersal of the test microorganism (p < 0.01). Hence, the results suggested that synergistic action of cuminaldehyde and tobramycin could be applied for the efficient management of microbial biofilm.


Assuntos
Infecções por Pseudomonas , Tobramicina , Humanos , Tobramicina/farmacologia , Antibacterianos/farmacologia , Pseudomonas aeruginosa , Biofilmes , Testes de Sensibilidade Microbiana , Sinergismo Farmacológico
15.
Appl Biochem Biotechnol ; 195(5): 3229-3256, 2023 May.
Artigo em Inglês | MEDLINE | ID: mdl-36580259

RESUMO

Gram-positive and Gram-negative bacteria often develop biofilm through different mechanisms in promoting pathogenicity. Hence, the antibiofilm molecule needs to be examined separately on both organisms to manage the biofilm threat. Since the antibiofilm activity of piperine against Staphylococcus aureus was already reported; here, we aimed to examine the antibiofilm activity of it against Pseudomonas aeruginosa. P. aeruginosa is an opportunistic Gram-negative pathogen that can cause several healthcare-associated infections by exploiting biofilm. Several experiments like crystal violet assay, estimation of total protein, measurement of extracellular polymeric substance, and microscopic analysis confirmed that lower concentrations (8 and 16 µg/mL) of piperine could inhibit the microbial biofilm formation considerably. Besides, it could also reduce the secretion of virulence factors from P. aeruginosa. Further investigation showed that the cell surface hydrophobicity and microbial motility of the test organism got reduced under the influence of piperine. Piperine exposure was found to increase the accumulation of reactive oxygen species (ROS) that resulted in the inhibition of biofilm formation. Furthermore, the molecular simulation studies suggested that piperine could affect the quorum sensing network of P. aeruginosa. Towards this direction, we noticed that piperine treatment could decrease the expression of the quorum sensing gene (lasI) that resulted in the inhibition of biofilm formation. Besides biofilm inhibition, piperine was also found to disintegrate the pre-existing biofilm of P. aeruginosa without showing any antimicrobial property to the test organism. Thus, piperine could be used for the sustainable protection of public-healthcare by compromising the biofilm assembly of P. aeruginosa.


Assuntos
Pseudomonas aeruginosa , Percepção de Quorum , Pseudomonas aeruginosa/genética , Espécies Reativas de Oxigênio , Antibacterianos/farmacologia , Antibacterianos/química , Matriz Extracelular de Substâncias Poliméricas/metabolismo , Bactérias Gram-Negativas , Bactérias Gram-Positivas , Biofilmes , Fatores de Virulência/metabolismo , Interações Hidrofóbicas e Hidrofílicas
16.
Folia Microbiol (Praha) ; 68(5): 801-811, 2023 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-37097592

RESUMO

Microorganisms embedded within an extracellular polymeric matrix are known as biofilm. The extensive use of antibiotics to overcome the biofilm-linked challenges has led to the emergence of multidrug-resistant strains. Staphylococcus aureus is one such nosocomial pathogen that is known to cause biofilm-linked infections. Thus, novel strategies have been adopted in this study to inhibit the biofilm formation of S. aureus. Two natural compounds, namely, 1,4-naphthoquinone (a quinone derivative) and tryptophan (aromatic amino acid), have been chosen as they could independently show efficient antibiofilm activity. To enhance the antibiofilm potential, the two compounds were combined and tested against the same organism. Several experiments like crystal violet (CV) assay, protein estimation, extracellular polymeric substance (EPS) extraction, and estimation of metabolic activity confirmed that the combination of the two compounds could significantly inhibit the biofilm formation of S. aureus. To comprehend the underlying mechanism, efforts were further directed to understand whether the two compounds could inhibit biofilm formation by compromising the cell surface hydrophobicity of the bacteria. The results revealed that the cell surface hydrophobicity got reduced by ~ 49% when the compounds were applied together. Thus, the combinations could show enhanced antibiofilm activity by attenuating cell surface hydrophobicity. Further studies revealed that the selected concentrations of the compounds could disintegrate (~ 70%) the pre-existing biofilm of the test bacteria without showing any antimicrobial activity. Hence, the combined application of tryptophan and 1,4-naphthoquinone could be used to inhibit the biofilm threats of S. aureus.


Assuntos
Infecções Estafilocócicas , Staphylococcus aureus , Humanos , Triptofano/farmacologia , Matriz Extracelular de Substâncias Poliméricas , Antibacterianos/farmacologia , Infecções Estafilocócicas/tratamento farmacológico , Infecções Estafilocócicas/microbiologia , Biofilmes , Testes de Sensibilidade Microbiana
17.
Folia Microbiol (Praha) ; 68(6): 843-854, 2023 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-37142893

RESUMO

1, 4-naphthoquinone, a plant-based quinone derivative, has gained much attention for its effectiveness against several biofilm-linked diseases. The biofilm inhibitory effect of 1, 4-naphthoquinone against Staphylococcus aureus has already been reported in our previous study. We observed that the extracellular DNA (eDNA) could play an important role in holding the structural integrity of the biofilm. Hence, in this study, efforts have been directed to examine the possible interactions between 1, 4-naphthoquinone and DNA. An in silico analysis indicated that 1, 4-naphthoquinone could interact with DNA through intercalation. To validate the same, UV-Vis spectrophotometric analysis was performed in which a hypochromic shift was observed when the said molecule was titrated with calf-thymus DNA (CT-DNA). Thermal denaturation studies revealed a change of 8℃ in the melting temperature (Tm) of CT-DNA when complexed with 1, 4-naphthoquinone. The isothermal calorimetric titration (ITC) assay revealed a spontaneous intercalation between CT-DNA and 1, 4-naphthoquinone with a binding constant of 0.95 ± 0.12 × 108. Furthermore, DNA was run through an agarose gel electrophoresis with a fixed concentration of ethidium bromide and increasing concentrations of 1, 4-naphthoquinone. The result showed that the intensity of ethidium bromide-stained DNA got reduced concomitantly with the gradual increase of 1, 4-naphthoquinone suggesting its intercalating nature. To gain further confidence, the pre-existing biofilm was challenged with ethidium bromide wherein we observed that it could also show biofilm disintegration. Therefore, the results suggested that 1, 4-naphthoquinone could exhibit disintegration of the pre-existing biofilm of Staphylococcus aureus through eDNA intercalation.


Assuntos
Naftoquinonas , Infecções Estafilocócicas , Humanos , Staphylococcus aureus/genética , Etídio/farmacologia , Naftoquinonas/farmacologia , DNA/farmacologia , Biofilmes
18.
Artigo em Inglês | MEDLINE | ID: mdl-38133854

RESUMO

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.

19.
Microsc Res Tech ; 85(7): 2628-2650, 2022 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-35411986

RESUMO

Pollen grains of 58 wetland species of angiosperms belonging to 17 families, growing in different wetlands of Sub-Himalayan biodiversity hotspots, Northern Bengal and lower Gangetic plains, India were investigated during a 4-year span of 2015-2019. Freshly collected anthers were processed through the acetolysis method and the obtained pollen grains were accurately studied under LM and SEM and properly photographed. Different attributes viz. shape, size, aperture type and exine ornamentation of pollen grains served as consistent features were used for authentic identification of the studied species. To study the phylogeny of pollen grains, characters like polarity, symmetry, aperture and exine sculpturing were found most significant for species segregation. Investigated pollen grains were mostly prolate-spheroidal, spheroidal or triangular, rarely prolate or elongated, exine psilate, granulate, reticulate or microechinate type. Most of the observable aperture of pollen grains of studied species were colporate, colpate and porate, while ulcerate and inaperturate grains were also observed in the studied aquatic members of the families Cyperaceae and Potamogetonaceae. All the gathered data were further statistically analyzed through ANOVA, PCA and Pearson Correlation Matrix to understand the interrelationship among the species. The permanent slides of identified pollen grains were deposited at the herbarium of North Bengal University [NBU] for future references. HIGHLIGHTS: Study of Pollen morphology of 58 wetland species under light & scanning electron microscopy. Unique apertures and exine ornamentation used to understand taxonomic evaluation of studied species. Pollen character supports interrelationship among the studied species. Relationship between exine thickness and the habitat of the wetland plants.


Assuntos
Magnoliopsida , Fosmet , Elétrons , Humanos , Magnoliopsida/anatomia & histologia , Microscopia Eletrônica de Varredura , Pólen/ultraestrutura , Áreas Alagadas
20.
3 Biotech ; 11(11): 485, 2021 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-34790509

RESUMO

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.

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