Anti-adhering Property Study of Green Nanotechnology Based …
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2.3 Determination of Minimum Inhibitory Concentration
(MIC)
The MIC of antibiotics (Vancomycin and Gentamycin) against S. aureus ATCC
25923 was determined by CLSI (2019) guidelines while those of crude C. longa and
A. indica plant extracts and the synthesized green nanoparticles were determined by
Kirby–Bauer disk diffusion technique.
2.4 Modification of Chitin Flakes with Antibiotics and Green
Nanoparticles
Based on the MIC, chitin flakes were encrusted with antibiotics, crude extracts of C.
longa and A. indica and green nanoparticles at 1 mg–50 mg/ml concentration for a
certain period of time and then used for testing.
2.5 Effect of Green Nanoparticle on Biofilm Formation
S. aureus ATCC 25923 biofilm was developed on chitin flasks by standard protocol
(Anderl et al. 2000). The bacterial culture was inoculated into Tryptic Soy Broth
and incubated overnight aerobically at 37 °C for 18 h. The overnight culture with a
concentration of 1 × 10
6 CFU/ml was inoculated into sterile TSB with normal and
encrusted chitin flakes and incubated at 37 °C for 24 h, 48 h, 72 h, and 96 h. The
chitin flakes were then washed with 0.9% saline by agitation at 180 rpm for 2 min to
remove non-adherent cells. Viable counts were determined by inoculating the biofilm
cells on TSA plates and incubated overnight at 37 °C (Gomes et al. 2012). SEM was
used to analyze the morphology of the biofilm structure on encrusted chitins and the
change in the chemical components was observed by FTIR.
3 Result
3.1 UV Analysis of Synthesized Green Nanoparticles
Ultraviolet–visible (UV–vis) absorption spectroscopic analysis showed the formation of silver nanoparticles from C. longa and A. indica extracts at a range of 300–
800 nm with a prominent peak at 429 and 420 nm (Fig. 1). Kumar et al. (2014) had
reported similar peak of silver nanoparticles was around 420 nm, and in the present
study, it was centered at 420–430 nm.
487
2.3 Determination of Minimum Inhibitory Concentration
(MIC)
The MIC of antibiotics (Vancomycin and Gentamycin) against S. aureus ATCC
25923 was determined by CLSI (2019) guidelines while those of crude C. longa and
A. indica plant extracts and the synthesized green nanoparticles were determined by
Kirby–Bauer disk diffusion technique.
2.4 Modification of Chitin Flakes with Antibiotics and Green
Nanoparticles
Based on the MIC, chitin flakes were encrusted with antibiotics, crude extracts of C.
longa and A. indica and green nanoparticles at 1 mg–50 mg/ml concentration for a
certain period of time and then used for testing.
2.5 Effect of Green Nanoparticle on Biofilm Formation
S. aureus ATCC 25923 biofilm was developed on chitin flasks by standard protocol
(Anderl et al. 2000). The bacterial culture was inoculated into Tryptic Soy Broth
and incubated overnight aerobically at 37 °C for 18 h. The overnight culture with a
concentration of 1 × 10
6 CFU/ml was inoculated into sterile TSB with normal and
encrusted chitin flakes and incubated at 37 °C for 24 h, 48 h, 72 h, and 96 h. The
chitin flakes were then washed with 0.9% saline by agitation at 180 rpm for 2 min to
remove non-adherent cells. Viable counts were determined by inoculating the biofilm
cells on TSA plates and incubated overnight at 37 °C (Gomes et al. 2012). SEM was
used to analyze the morphology of the biofilm structure on encrusted chitins and the
change in the chemical components was observed by FTIR.
3 Result
3.1 UV Analysis of Synthesized Green Nanoparticles
Ultraviolet–visible (UV–vis) absorption spectroscopic analysis showed the formation of silver nanoparticles from C. longa and A. indica extracts at a range of 300–
800 nm with a prominent peak at 429 and 420 nm (Fig. 1). Kumar et al. (2014) had
reported similar peak of silver nanoparticles was around 420 nm, and in the present
study, it was centered at 420–430 nm.