Concentration Dependent Effect of Azadirachta indica (Neem) Seed Oil and Neem Bark extract on Planktonic and Established Biofilm Growth of Pseudomonas aeruginosa and Staphylococcus aureus

Azadirachta indica Juss (Neem) is well documented for its antimicrobial activity. The effect of varying concentrations (0.1 to 50% v/v) of Azadirachta indica derived neem seed oil (NSO), neem seed oil with tween 20 and neem bark extract was evaluated on planktonic, biofilm formation and mature biofi...

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Main Authors: Ravish Katiyar, Ekta Khare, Shilpa Deshpande Kaistha
Format: Article
Language:English
Published: Journal of Pure and Applied Microbiology 2023-09-01
Series:Journal of Pure and Applied Microbiology
Subjects:
Online Access:https://microbiologyjournal.org/concentration-dependent-effect-of-azadirachta-indica-neem-seed-oil-and-neem-bark-extract-on-planktonic-and-established-biofilm-growth-of-pseudomonas-aeruginosa-and-staphylococcus-aureus/
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author Ravish Katiyar
Ekta Khare
Shilpa Deshpande Kaistha
author_facet Ravish Katiyar
Ekta Khare
Shilpa Deshpande Kaistha
author_sort Ravish Katiyar
collection DOAJ
description Azadirachta indica Juss (Neem) is well documented for its antimicrobial activity. The effect of varying concentrations (0.1 to 50% v/v) of Azadirachta indica derived neem seed oil (NSO), neem seed oil with tween 20 and neem bark extract was evaluated on planktonic, biofilm formation and mature biofilms of multiple drug resistant Pseudomonas aeruginosa ATCC 15442 and Staphylococcus aureus ATCC 25923 using the crystal violet assay and scanning electron microscopy. NSO showed antimicrobial activity at 25% v/v for P. aeruginosa but not S. aureus in zone of inhibition assay. Neem bark extract on the contrary showed antimicrobial activity against both the isolates at 50% v/v concentrations. Interestingly, in biofilm formation assay, low concentrations of NSO (3.5 to 0.2% v/v) induced biofilm formation while inhibition of both planktonic and biofilm was seen in concentration dependent manner from 12.5% v/v onwards. Complex of NSO and tween in comparison of NSO alone caused low induction in S.aureus biofilm formation, while inhibiting biofilm formation of P. aeruginosa at all the concentrations. In biofilm eradication assay, NSO induced biofilm of both P. aeruginosa (50 to 0.1%v/v) and S. aureus (50 to 3.13%v/v). Eradication effect of neem bark extract was found on P. aeruginosa biofilm in a dose dependent fashion from 50 to 20% v/v followed by 0.2 to 0.1%v/v concentration respectively. S. aureus biofilm were eradicated at 50 to 25%v/v concentrations. At low concentrations, both the neem derivatives induced biofilm mediated growth of the pathogenic organisms. The data also indicate that neem seed oil was more effective against Gram negative P. aeruginosa while neem bark extract was effective against Gram positive S. aureus. This study highlights the crucial but variable effects of concentration dependent effect of phytochemicals and their composition on biofilm induction as well as eradication, the primary growth form in clinical settings. This challenges the notion that all herbal products are safe as antimicrobial activities differ as per microbial growth modes. Hence, concentration dependent effect of medicinal plant derived products requires thorough investigation prior to their use as antimicrobial agents.
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spelling doaj.art-b3d2f9ddce064b279ca26294fcf0ba3d2023-09-15T15:30:49ZengJournal of Pure and Applied MicrobiologyJournal of Pure and Applied Microbiology0973-75102581-690X2023-09-0117316691678https://doi.org/10.22207/JPAM.17.3.30Concentration Dependent Effect of Azadirachta indica (Neem) Seed Oil and Neem Bark extract on Planktonic and Established Biofilm Growth of Pseudomonas aeruginosa and Staphylococcus aureusRavish Katiyarhttps://orcid.org/0009-0007-9708-566XEkta Kharehttps://orcid.org/0000-0002-0531-8267Shilpa Deshpande Kaisthahttps://orcid.org/0000-0002-2196-7224Azadirachta indica Juss (Neem) is well documented for its antimicrobial activity. The effect of varying concentrations (0.1 to 50% v/v) of Azadirachta indica derived neem seed oil (NSO), neem seed oil with tween 20 and neem bark extract was evaluated on planktonic, biofilm formation and mature biofilms of multiple drug resistant Pseudomonas aeruginosa ATCC 15442 and Staphylococcus aureus ATCC 25923 using the crystal violet assay and scanning electron microscopy. NSO showed antimicrobial activity at 25% v/v for P. aeruginosa but not S. aureus in zone of inhibition assay. Neem bark extract on the contrary showed antimicrobial activity against both the isolates at 50% v/v concentrations. Interestingly, in biofilm formation assay, low concentrations of NSO (3.5 to 0.2% v/v) induced biofilm formation while inhibition of both planktonic and biofilm was seen in concentration dependent manner from 12.5% v/v onwards. Complex of NSO and tween in comparison of NSO alone caused low induction in S.aureus biofilm formation, while inhibiting biofilm formation of P. aeruginosa at all the concentrations. In biofilm eradication assay, NSO induced biofilm of both P. aeruginosa (50 to 0.1%v/v) and S. aureus (50 to 3.13%v/v). Eradication effect of neem bark extract was found on P. aeruginosa biofilm in a dose dependent fashion from 50 to 20% v/v followed by 0.2 to 0.1%v/v concentration respectively. S. aureus biofilm were eradicated at 50 to 25%v/v concentrations. At low concentrations, both the neem derivatives induced biofilm mediated growth of the pathogenic organisms. The data also indicate that neem seed oil was more effective against Gram negative P. aeruginosa while neem bark extract was effective against Gram positive S. aureus. This study highlights the crucial but variable effects of concentration dependent effect of phytochemicals and their composition on biofilm induction as well as eradication, the primary growth form in clinical settings. This challenges the notion that all herbal products are safe as antimicrobial activities differ as per microbial growth modes. Hence, concentration dependent effect of medicinal plant derived products requires thorough investigation prior to their use as antimicrobial agents.https://microbiologyjournal.org/concentration-dependent-effect-of-azadirachta-indica-neem-seed-oil-and-neem-bark-extract-on-planktonic-and-established-biofilm-growth-of-pseudomonas-aeruginosa-and-staphylococcus-aureus/neem seed oilneem barkpseudomonas aeruginosastaphylococcus aureusbiofilm
spellingShingle Ravish Katiyar
Ekta Khare
Shilpa Deshpande Kaistha
Concentration Dependent Effect of Azadirachta indica (Neem) Seed Oil and Neem Bark extract on Planktonic and Established Biofilm Growth of Pseudomonas aeruginosa and Staphylococcus aureus
Journal of Pure and Applied Microbiology
neem seed oil
neem bark
pseudomonas aeruginosa
staphylococcus aureus
biofilm
title Concentration Dependent Effect of Azadirachta indica (Neem) Seed Oil and Neem Bark extract on Planktonic and Established Biofilm Growth of Pseudomonas aeruginosa and Staphylococcus aureus
title_full Concentration Dependent Effect of Azadirachta indica (Neem) Seed Oil and Neem Bark extract on Planktonic and Established Biofilm Growth of Pseudomonas aeruginosa and Staphylococcus aureus
title_fullStr Concentration Dependent Effect of Azadirachta indica (Neem) Seed Oil and Neem Bark extract on Planktonic and Established Biofilm Growth of Pseudomonas aeruginosa and Staphylococcus aureus
title_full_unstemmed Concentration Dependent Effect of Azadirachta indica (Neem) Seed Oil and Neem Bark extract on Planktonic and Established Biofilm Growth of Pseudomonas aeruginosa and Staphylococcus aureus
title_short Concentration Dependent Effect of Azadirachta indica (Neem) Seed Oil and Neem Bark extract on Planktonic and Established Biofilm Growth of Pseudomonas aeruginosa and Staphylococcus aureus
title_sort concentration dependent effect of azadirachta indica neem seed oil and neem bark extract on planktonic and established biofilm growth of pseudomonas aeruginosa and staphylococcus aureus
topic neem seed oil
neem bark
pseudomonas aeruginosa
staphylococcus aureus
biofilm
url https://microbiologyjournal.org/concentration-dependent-effect-of-azadirachta-indica-neem-seed-oil-and-neem-bark-extract-on-planktonic-and-established-biofilm-growth-of-pseudomonas-aeruginosa-and-staphylococcus-aureus/
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AT ektakhare concentrationdependenteffectofazadirachtaindicaneemseedoilandneembarkextractonplanktonicandestablishedbiofilmgrowthofpseudomonasaeruginosaandstaphylococcusaureus
AT shilpadeshpandekaistha concentrationdependenteffectofazadirachtaindicaneemseedoilandneembarkextractonplanktonicandestablishedbiofilmgrowthofpseudomonasaeruginosaandstaphylococcusaureus