Abstract
To improve the efficacy of existing classes of antibiotics (ciprofloxacin), allow dose reduction, and minimize related toxicity, this study was executed because new target-oriented livestock antimicrobials are greatly needed to battle infections caused by multidrug-resistant (MDR) strains. The present study aims to green synthesize a biocompatible nanohybrid of ciprofloxacin (CIP)-Ag/TiO2/chitosan (CS). Silver and titanium nanoparticles were green synthesized using Moringa concanensis leaves extract. The incorporation of silver (Ag) nanoparticles onto the surface of titanium oxide nanoparticles (TiO2NPs) was done by the wet chemical impregnation method, while the encapsulation of chitosan (CS) around Ag/TiO2 conjugated with ciprofloxacin (CIP) was done by the ionic gelation method. The synthesized nanohybrid (CIP-Ag/TiO2/CS) was characterized using standard techniques. The antibacterial potential, killing kinetics, cytotoxicity, drug release profile, and minimum inhibitory concentration (MIC) were determined. Field emission scanning electron microscopy (FESEM) and transmission electron microscopy (TEM) revealed spherical agglomerated nanoparticles (NPs) of Ag/TiO2 with particle sizes of 47–75 nm, and those of the CIP-Ag/TiO2/CS nanohybrid were in range of 20–80 nm. X-ray diffractometer (XRD) patterns of the hetero system transmitted diffraction peaks of anatase phase of TiO2 and centered cubic metallic Ag crystals. Fourier Transform Infrared spectroscopy (FTIR) confirmed the Ti-O-Ag linkage in the nanohybrid. The zeta potential of CIP-Ag/TiO2/CS nanohybrid was found (67.45 ± 1.8 mV), suggesting stable nanodispersion. The MIC of CIP-Ag/TiO2/CS was 0.0512 μg/mL, which is much lower than the reference value recorded by the global CLSI system (Clinical Laboratory Standards Institute). The CIP-Ag/TiO2/CS nanohybrid was found to be effective against mastitis causing MDR E. coli; killing kinetics showed an excellent reduction of E. coli cells at 6 h of treatment. Flow cytometry further confirmed antibacterial potential by computing 67.87% late apoptosis feature at 6 h of treatment; antibiotic release kinetic revealed a sustained release of CIP. FESEM and TEM confirmed the structural damages in MDR E. coli (multidrug-resistant Escherichia coli). The CIP-Ag/TiO2/CS nanohybrid was found to be biocompatible, as more than 93.08% of bovine mammary gland epithelial cells remained viable. The results provide the biological backing for the development of nanohybrid antibiotics at a lower MIC value to treat infectious diseases of cattle and improve the efficacy of existing classes of antibiotics by conjugation with nanoparticles.
Highlights
The use of antibiotics in the treatment of animal infection and the enhancement of animal health are the main driving forces behind the development of antimicrobial resistance in animal husbandry [1,2]
Considering the urgent need for new clinical interventions to control the global issue of antimicrobial resistance (AMR), a unique ciprofloxacin loaded-TiO2/Ag/CS nanohybrid was synthesized by the green approach using M. concanensis leaves extract
The results showed that E. coli was found highly resistant to the current range of antibiotics, raising concern to find the solution for the emerging resistant Gram-negative rods of mastitis-suffered cattle, as shown in the Figure 1
Summary
The use of antibiotics in the treatment of animal infection and the enhancement of animal health are the main driving forces behind the development of antimicrobial resistance in animal husbandry [1,2]. Traditional antibiotics have been less effective in the treatment of infectious diseases in dairy cattle In this context, there is an urgent need for the production of biocompatible antibacterial formulations that could regulate bacterial growth by means of improved and efficient mechanisms [4]. The plant-mediated green synthesis of silver nanoparticles has grown into a unique and novel field of nanotechnology It has gained importance because of its ecofriendly and cost efficiency with lower toxicity as compared to chemical methods [18]. Considering the urgent need for new clinical interventions to control the global issue of antimicrobial resistance (AMR), a unique ciprofloxacin loaded-TiO2/Ag/CS nanohybrid was synthesized by the green approach using M. concanensis leaves extract. A ciprofloxacin loaded-TiO2/Ag/CS nanohybrid was tested for its efficacy against MDR strains of E. coli-causing mastitis in the cattle by various standard antimicrobial assays indicating the strong antibacterial potential of synthesized nanohybrids. A standard API20E test panel was used for the biochemical-based identification of E. coli [22]
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