Revisiting the Synergistic In Vitro Antimicrobial and Antibiofilm Potential of Chlorhexidine Gluconate and Cetrimide in Combination as an Antiseptic and Disinfectant Agent
Chlorhexidine and cetrimide are often used as antiseptics and disinfectants. While their individual activities are well-documented, their synergism has rarely been evaluated. Here, we attempted to evaluate the antimicrobial and antibiofilm effects of the combination of these two antimicrobial agents...
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2025-01-01
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| author | Diamond Jain Rimjhim Gupta Rashmi Mehta Pratheesh N. Prabhakaran Deva Kumari Kulpreet Bhui Deepa Murali |
| author_facet | Diamond Jain Rimjhim Gupta Rashmi Mehta Pratheesh N. Prabhakaran Deva Kumari Kulpreet Bhui Deepa Murali |
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| description | Chlorhexidine and cetrimide are often used as antiseptics and disinfectants. While their individual activities are well-documented, their synergism has rarely been evaluated. Here, we attempted to evaluate the antimicrobial and antibiofilm effects of the combination of these two antimicrobial agents against two environment isolates, viz., <i>P. aeruginosa</i> and <i>S. aureus</i>. The synergism was assayed by determining the fractional inhibitory concentrations, while the antibiofilm effects were determined using crystal violet staining and the resazurin assay. Further, the effects on the biofilms were visualized using brightfield and confocal laser scanning microscopy. Our results show that the combination of these antimicrobials resulted in synergistic inhibition of <i>P. aeruginosa</i> growth. When tested at concentrations below the individual MICs (one-quarter of the MICs), the combination was able to significantly reduce the adherence of <i>S. aureus</i> biofilms to a polystyrene surface, while no effect was observed for <i>P. aeruginosa</i>. The combination was also able to significantly reduce the viability of pre-formed biofilms of both bacteria, thereby showing its antibiofilm potential. Next, we evaluated the performance of this combination against a wide array of micro-organisms. This fixed-dose combination formulation exhibited a significant reduction in the viability of an array of clinically relevant micro-organisms, including ESKAPE pathogens, <i>Mycobacterium</i> sp., MRSA, <i>Leptospira</i>, <i>Candida</i> sp., norovirus and adenovirus. Overall, it can be inferred that the combination of chlorhexidine and cetrimide is a potential biocide that continues to be relevant for use in antisepsis and disinfection against infection-causing pathogens. |
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| language | English |
| publishDate | 2025-01-01 |
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| series | Microbiology Research |
| spelling | doaj-art-2b53116e9444477799e37ef1127694082025-01-24T13:41:44ZengMDPI AGMicrobiology Research2036-74812025-01-011611610.3390/microbiolres16010016Revisiting the Synergistic In Vitro Antimicrobial and Antibiofilm Potential of Chlorhexidine Gluconate and Cetrimide in Combination as an Antiseptic and Disinfectant AgentDiamond Jain0Rimjhim Gupta1Rashmi Mehta2Pratheesh N. Prabhakaran3Deva Kumari4Kulpreet Bhui5Deepa Murali6Savlon Swasth India Mission, ITC Life Sciences and Technology Centre, Bangalore 560057, IndiaSavlon Swasth India Mission, ITC Life Sciences and Technology Centre, Bangalore 560057, IndiaSavlon Swasth India Mission, ITC Life Sciences and Technology Centre, Bangalore 560057, IndiaSavlon Swasth India Mission, ITC Life Sciences and Technology Centre, Bangalore 560057, IndiaSavlon Swasth India Mission, ITC Life Sciences and Technology Centre, Bangalore 560057, IndiaSavlon Swasth India Mission, ITC Life Sciences and Technology Centre, Bangalore 560057, IndiaSavlon Swasth India Mission, ITC Life Sciences and Technology Centre, Bangalore 560057, IndiaChlorhexidine and cetrimide are often used as antiseptics and disinfectants. While their individual activities are well-documented, their synergism has rarely been evaluated. Here, we attempted to evaluate the antimicrobial and antibiofilm effects of the combination of these two antimicrobial agents against two environment isolates, viz., <i>P. aeruginosa</i> and <i>S. aureus</i>. The synergism was assayed by determining the fractional inhibitory concentrations, while the antibiofilm effects were determined using crystal violet staining and the resazurin assay. Further, the effects on the biofilms were visualized using brightfield and confocal laser scanning microscopy. Our results show that the combination of these antimicrobials resulted in synergistic inhibition of <i>P. aeruginosa</i> growth. When tested at concentrations below the individual MICs (one-quarter of the MICs), the combination was able to significantly reduce the adherence of <i>S. aureus</i> biofilms to a polystyrene surface, while no effect was observed for <i>P. aeruginosa</i>. The combination was also able to significantly reduce the viability of pre-formed biofilms of both bacteria, thereby showing its antibiofilm potential. Next, we evaluated the performance of this combination against a wide array of micro-organisms. This fixed-dose combination formulation exhibited a significant reduction in the viability of an array of clinically relevant micro-organisms, including ESKAPE pathogens, <i>Mycobacterium</i> sp., MRSA, <i>Leptospira</i>, <i>Candida</i> sp., norovirus and adenovirus. Overall, it can be inferred that the combination of chlorhexidine and cetrimide is a potential biocide that continues to be relevant for use in antisepsis and disinfection against infection-causing pathogens.https://www.mdpi.com/2036-7481/16/1/16antisepsisantimicrobial resistancedisinfection<i>Leptospira</i>chlorhexidinecetrimide |
| spellingShingle | Diamond Jain Rimjhim Gupta Rashmi Mehta Pratheesh N. Prabhakaran Deva Kumari Kulpreet Bhui Deepa Murali Revisiting the Synergistic In Vitro Antimicrobial and Antibiofilm Potential of Chlorhexidine Gluconate and Cetrimide in Combination as an Antiseptic and Disinfectant Agent Microbiology Research antisepsis antimicrobial resistance disinfection <i>Leptospira</i> chlorhexidine cetrimide |
| title | Revisiting the Synergistic In Vitro Antimicrobial and Antibiofilm Potential of Chlorhexidine Gluconate and Cetrimide in Combination as an Antiseptic and Disinfectant Agent |
| title_full | Revisiting the Synergistic In Vitro Antimicrobial and Antibiofilm Potential of Chlorhexidine Gluconate and Cetrimide in Combination as an Antiseptic and Disinfectant Agent |
| title_fullStr | Revisiting the Synergistic In Vitro Antimicrobial and Antibiofilm Potential of Chlorhexidine Gluconate and Cetrimide in Combination as an Antiseptic and Disinfectant Agent |
| title_full_unstemmed | Revisiting the Synergistic In Vitro Antimicrobial and Antibiofilm Potential of Chlorhexidine Gluconate and Cetrimide in Combination as an Antiseptic and Disinfectant Agent |
| title_short | Revisiting the Synergistic In Vitro Antimicrobial and Antibiofilm Potential of Chlorhexidine Gluconate and Cetrimide in Combination as an Antiseptic and Disinfectant Agent |
| title_sort | revisiting the synergistic in vitro antimicrobial and antibiofilm potential of chlorhexidine gluconate and cetrimide in combination as an antiseptic and disinfectant agent |
| topic | antisepsis antimicrobial resistance disinfection <i>Leptospira</i> chlorhexidine cetrimide |
| url | https://www.mdpi.com/2036-7481/16/1/16 |
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