In vitro and in vivo biofilm inhibitory efficacy of geraniol-cefotaxime combination against Staphylococcus spp.

Kannappan, Arunachalam; Balasubramaniam, Boopathi; Ranjitha, Rajendhran; Ramanathan, S.; Packiavathy, Issac Abraham Sybiya Vasantha; Balamurugan, Krishnaswamy; Pandian, Shunmugiah Karutha; Ravi, Arumugam Veera

doi:10.1016/j.fct.2019.01.008

Cited by 47 publications

(27 citation statements)

References 38 publications

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“…The Z-stack analysis was done with the Zen 2009 software (Carl Zeiss, Germany). In addition, biofilm biovolume and average thickness of the biofilm formed in control and treated samples (z-stack images) were analyzed by COMSTAT software (kindly gifted by Dr. Claus Sternberg, DTU Systems Biology, Technical University of Denmark; Kannappan et al, 2019).…”

Section: Confocal Laser Scanning Microscope Analysismentioning

confidence: 99%

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AHL-Lactonase Producing Psychrobacter sp. From Palk Bay Sediment Mitigates Quorum Sensing-Mediated Virulence Production in Gram Negative Bacterial Pathogens

et al. 2021

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Quorum sensing (QS) is a signaling mechanism governed by bacteria used to converse at inter- and intra-species levels through small self-produced chemicals called N-acylhomoserine lactones (AHLs). Through QS, bacteria regulate and organize the virulence factors’ production, including biofilm formation. AHLs can be degraded by an action called quorum quenching (QQ) and hence QQ strategy can effectively be employed to combat biofilm-associated bacterial pathogenesis. The present study aimed to identify novel bacterial species with QQ potential. Screening of Palk Bay marine sediment bacteria for QQ activity ended up with the identification of marine bacterial isolate 28 (MSB-28), which exhibited a profound QQ activity against QS biomarker strain Chromobacterium violaceum ATCC 12472. The isolate MSB-28 was identified as Psychrobacter sp. through 16S-rRNA sequencing. Psychrobacter sp. also demonstrated a pronounced activity in controlling the biofilm formation in different bacteria and biofilm-associated virulence factors’ production in P. aeruginosa PAO1. Solvent extraction, heat inactivation, and proteinase K treatment assays clearly evidence the enzymatic nature of the bioactive lead. Furthermore, AHL’s lactone ring cleavage was confirmed with experiments including ring closure assay and chromatographic analysis, and thus the AHL-lactonase enzyme production in Psychrobacter sp. To conclude, this is the first report stating the AHL-lactonase mediated QQ activity from marine sediment bacteria Psychrobacter sp. Future work deals with the characterization, purification, and mass cultivation of the purified protein and should pave the way to assessing the feasibility of the identified protein in controlling QS and biofilm-mediated multidrug resistant bacterial infections in mono or multi-species conditions.

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Section: Confocal Laser Scanning Microscope Analysismentioning

confidence: 99%

“…interferes with the biofilm formation of tested bacterial pathogens. COMSTAT software analysis was a quantitative method employed to analyze the biofilm formation of the tested pathogen using the raw LSM file (Kannappan et al, 2019).…”

Section: Microscopy Analysis -Clsm Observationmentioning

confidence: 99%

AHL-Lactonase Producing Psychrobacter sp. From Palk Bay Sediment Mitigates Quorum Sensing-Mediated Virulence Production in Gram Negative Bacterial Pathogens

et al. 2021

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“… 30 Results of the CRA assay showed that carvacrol reduced the slime synthesis of MRSA, and it well correlated with the previous reports, wherein the inhibitory potential of antibiofilm agents was explored on the slime synthesis of MRSA. 13 , 31 , 32 Further, EPS is a major component of the bacterial biofilm, and it contains polysaccharide proteins and nucleic acids. It supports the formation of the biofilm architecture and persistence of the bacterial biofilm.…”

Section: Discussionmentioning

confidence: 99%

Carvacrol Targets SarA and CrtM of Methicillin-Resistant Staphylococcus aureus to Mitigate Biofilm Formation and Staphyloxanthin Synthesis: An In Vitro and In Vivo Approach

et al. 2020

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Carvacrol is an essential oil traditionally used in culinary processes as spice due to its aromatic nature and also known for various biological activities. In the present study, the antivirulence efficacy of carvacrol against methicillin-resistant Staphylococcus aureus (MRSA) is explored. MRSA is an opportunistic pathogen capable of causing various superficial and systemic infections in humans. Biofilm formation and virulence factors of MRSA are responsible for its pathogenesis and resistance. Hence, the aim of this study was to explore the antibiofilm and antivirulence efficacy of carvacrol against MRSA. Carvacrol at 75 μg/mL inhibited MRSA biofilm by 93%, and it also decreased the biofilm formation on polystyrene and glass surfaces. Further, microscopic analyses revealed the reduction in microcolony formation and collapsed structure of biofilm upon carvacrol treatment. The growth curve analysis and the Alamar blue assay showed the nonfatal effect of carvacrol on MRSA. Further, carvacrol significantly reduced the production of MRSA biofilm-associated slime and extracellular polysaccharide. In addition, carvacrol strongly inhibited the antioxidant pigment staphyloxanthin and its intermediates' synthesis in MRSA. Inhibition of biofilm and staphyloxanthin by carvacrol enhanced the susceptibility of MRSA to oxidants and healthy human blood. Quantitative polymerase chain reaction (qPCR) analysis unveiled the downregulation of sarA-mediated biofilm gene expression and staphyloxanthin-associated crtM gene expression. The sarA-dependent antibiofilm potential of carvacrol was validated using S. aureus Newman wild-type and isogenic ΔsarA strains. In silico molecular docking analysis showed the high binding efficacy of carvacrol with staphylococcal accessory regulator A (SarA) and 4,4′diapophytoene synthase (CrtM) when compared to positive controls. Furthermore, the in vivo efficacy of carvacrol against MRSA infection was demonstrated using the model organism Galleria mellonella. The results revealed the nontoxic nature of carvacrol to the larvae and the rescuing potential of carvacrol against MRSA infection. Finally, the current study reveals the potential of carvacrol in inhibiting the biofilm formation and staphyloxanthin synthesis of MRSA by targeting the global regulator SarA and a novel antivirulence target CrtM.

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“…The biofilm‐forming capacity assay was performed by following the method of Kannappan et al . (2019). Briefly, the cultures of bacterial strains were diluted (1 : 100) and incubated in 48‐well microtiter plates at 4°C for 7 days.…”

Section: Methodsmentioning

confidence: 99%

Characterization of co‐culture of Aeromonas and Pseudomonas bacterial biofilm and spoilage potential on refrigerated grass carp ( Ctenopharyngodon idellu s)

Zhang

Zhu

Jatt

et al. 2020

Lett Appl Microbiol

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Aeromonas and Pseudomonas are important bacterial species involved in spoilage of refrigerated freshwater fish. In this study, 10 Aeromonas and seven Pseudomonas bacterial strains were isolated from spoiled grass carp and identified. Twelve of seventeen bacterial strains showed high potential of biofilm formation and 14 of 17 can produce extracellular protease. In order to explore the spoilage capacity of dual‐species, the sterile grass carp fillets were inoculated with mono‐ and dual‐species of Aeromonas salmonicida and Pseudomonas azotoformans strains. The results revealed significantly higher levels of the total viable count and total volatile basic nitrogen in dual‐species as compared to mono‐species from day 6. The higher contents of histamine, cadaverine and serious degradation in muscles tissue were also observed in dual‐species after 10 days of storage. Results of in vitro experiments showed that the co‐culture of A. salmonicida and P. azotoformans significantly increased the bacterial maximum growth rate, promoted the biofilm formation and improved the spoilage capacity of bacterial strains. This study has revealed that the co‐culture of Aeromonas and Pseudomonas bacterial strains accelerated spoilage process of grass carp and increased biofilm formation. It indicates that the mixed‐cultures of spoilage micro‐organisms pose a huge threat to food industry.

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In vitro and in vivo biofilm inhibitory efficacy of geraniol-cefotaxime combination against Staphylococcus spp.

Cited by 47 publications

References 38 publications

AHL-Lactonase Producing Psychrobacter sp. From Palk Bay Sediment Mitigates Quorum Sensing-Mediated Virulence Production in Gram Negative Bacterial Pathogens

AHL-Lactonase Producing Psychrobacter sp. From Palk Bay Sediment Mitigates Quorum Sensing-Mediated Virulence Production in Gram Negative Bacterial Pathogens

Carvacrol Targets SarA and CrtM of Methicillin-Resistant Staphylococcus aureus to Mitigate Biofilm Formation and Staphyloxanthin Synthesis: An In Vitro and In Vivo Approach

Characterization of co‐culture of Aeromonas and Pseudomonas bacterial biofilm and spoilage potential on refrigerated grass carp ( Ctenopharyngodon idellu s)

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