Browsing by Author "Bharavi, K(MAJOR)"
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ThesisItem Open Access SYNTHESIS AND EVALUATION OF ANTIBACTERIAL ACTIVITY OF ENROFLOXACIN CONJUGATED NANO ZINC OXIDE(SRI VENKATESWARA VETERINARY UNIVERSITY TIRUPATI - 517 502. (A.P.) INDIA, 2018-05) Mulla Hussain Basha; Bharavi, K(MAJOR); Srinivasa Rao, G; Annapurna, PABSTRACT : Antimicrobial resistance is a major concern in veterinary medicine. In this study, the successful conjugation of enrofloxacin with amine functionalized zinc oxide nanoparticles (ZNP) was described and its antibacterial activity was evaluated against standard MTCC cultures and clinical isolates. ZNP were synthesized using microwave-assisted method using zinc acetate dihydrate. 3-aminopropyltriethoxysilane (3-APTES) was used to amine functionalize ZNP using co-condensation technique. Enrofloxacin was conjugated with amine functionalized ZNP using 1-Ethyl-3-(3-dimethylaminopropyl) carbodiimide (EDC) and N-hydroxysuccinimide (NHS). ZNP, enrofloxacin and enrofloxacin conjugated zinc oxide nanoparticles (ECZN) the conjugate were analyzed by using various techniques like UV-Vis spectrophotometery, Dynamic light scattering (DLS),Transmission electron microscopy (TEM), and Fourier-transform infrared spectroscopy (FTIR) analysis to assess the nano size and conjugation. The antibacterial activity of ECZN was evaluated using microtitre dilution method using standard MTCC cultures (S.aureus - MTCC 3160, S.pyogenes - MTCC 1927, S.typhimurium -MTCC 3224, E.faecalis - MTCC 9845, E.coli - MTCC 443, P.aeruginosa -MTCC 3542 and K.pneumoniae - MTCC 432) and clinical isolates (E.coli, Salmonella sps, and S. aureus). ZNP were spherical with uniform distribution and a diameter of 20 nm. The particles showed a characteristic peak at 380 nm with a hydrodynamic radius of 28.3 nm, zeta potential of -29.7 mV. The FTIR spectra of ZNP exhibited characteristic peaks at 3398.57 (3400) cm−1, 1553.79 (1632) cm−1. Successful amine functionalization of ZNP was confirmed by the observation of deep purple colour in ninhydrin test. The FTIR spectra of amine functionalized ZNP shows a characteristic peak at 3307.29 cm−1 (due to the presence of O–H and –N–H stretching of NH2 group); 2967.24 cm−1 (due to the asymmetric C–H stretching of the CH2 group of APTES); 1626.18 cm−1 and 1461.78 cm−1 (due to NH2 scissoring of primary amine). The conjugation of enrofloxacin with amine functionalized ZNP (efficacy: 1.18 mg%) was confirmed by UV-Vis spectra and FTIR Native enrofloxacin exhibited two characteristic peaks at 280 nm and 321 nm. After conjugation with ZNP, the second peak slightly shifted to 375 nm while the first peak remained the same. In the FTIR spectra ECZN showed peaks obtained in the region of 1400–1550 cm−1 which were similar to enrofloxacin peaks, while 896.47 cm−1 signified Zn– O stretching. The MIC (μg mL-1) of enrofloxacin was significantly (P<0.05) reduced against respective MTCC culture in combination with ZNP (enrofloxacin in native form vs enrofloxacin in ECZN) (S.aureus – 0.106 vs 0.0637*; S.pyogenes – 0.065 vs 0.0245*; S.typhimurium – 0.032 vs 0.0051*; E.faecalis – 0.021 vs 0.0083*; E.coli – 0.047 vs 0.0187*; P.aeruginosa – 2.611 vs 0.1444*; K.pneumoniae – 0.0.65 vs 0.0944*). Against clinical isolates, the MIC of native vs ECZN (actual enrofloxacin concentration) were comparable (E.coli – 0.0975 vs 0.083; Salmonella sps – 0.079 vs 0.069; S.auerus – 0.111 vs 0.086). However, the MIC of ECZN against both standard cultures and clinical isolates was significantly (P<0.05) lower than ZNP. In conclusion, enrofloxacin could be successfully conjugated with amine functionalized zinc oxide nanoparticles. The antibacterial efficacy of ZNP was improved in the combination with enrofloxacin against standard MTCC cultures and clinical isolates. In future, ways to improve the efficacy of enrofloxacin conjugation with zinc oxide nanoparticles and spectrum of activity could be explored.