Muller hinton agar

Castor oil (Amman Pharmaceutical industries CO, Jordan), activated charcoal (Research-Lab fine Chem industries, Mumbai, India), loperamide (Daehwa Pharmaceuticals, Republic of Korea), distilled water (Dallul Pharmaceuticals PLC, Addis Ababa, Ethiopia), methanol (Alpha Chemika, Mumbai, India), ciprofloxacin 5 μg/disc (Becton, Dickinson and Company, Sparks, USA), ceftazidime 30 μg/disc (Oxoid Ltd., Basingstoke, England), muller hinton agar (MHA) (HiMedia Laboratories Pvt. Ltd., India), muller hinton broth (MHB) (HiMedia Laboratories Pvt. Ltd., India) and nutrient agar (Micro master lab, India) were used during the study.

The chemicals used in the current experiments were: Fullerenes (C₆₀) and AgNPs were procured commercially from Sigma-Aldrich (St. Louis, MO, USA) of 99.99 % purity grade with no further purifications. The solvent used in this study was double-distilled water. The other chemicals, such as Muller Hinton Agar (MHA) and Potato Dextrose Agar (PDA) media, were purchased from Hi-Media. Moreover, DPPH, streptomycin sulfate, benzyl antibiotics, and doxorubicin were purchased from Sigma Aldrich. The cell line media such as Dulbecco’s Modified Eagle’s Medium (DMEM), Trypsin-EDTA and Fetal Bovine Serum (FBS) were purchased from GIBCOTM (Grand Island, NY, USA).

The bacterial isolate’s antibiotic sensitivity was assessed using a standard disc diffusion test following the CLSI guidelines [99 ,100 ]. To begin the assay, a bacterial starter culture was raised overnight from a single colony in NB at 28 °C and 200 rpm. The culture was then spread-plated on Muller Hinton agar (MHA; Himedia, Mumbai, India). After approximately 30 min of inoculum soaking, susceptibility discs containing various antibiotics (Himedia) were placed on the plates. These plates were subsequently incubated overnight in the dark at 28 °C. The extent of bacterial susceptibility to the antibiotics was determined by measuring the observed zone of inhibition, which represented the diameter of the region where bacterial growth was inhibited. A larger zone indicated greater susceptibility, while a smaller or absent zone indicated antibiotic resistance. The results were expressed as the means of the means obtained from three replicates, with each replicate stemming from three trials per antibiotic tested.

Chloroauric acid (HAuCl₄.3H₂O) was purchased from Sigma, USA. Zinc granulated metal (Zn) at a purity of 99.9% was purchased from VWR International, USA. Muller–Hinton agar and nutrient broth medium were purchased from HiMedia (India). The rat embryonic fibroblast (REF) cell line was kindly gifted from the Iraqi Center for Cancer and Medical Genetics Research (ICCMGR). RPMI-1640 medium was purchased from Gibco (USA). Fetal calf serum, streptomycin, penicillin, acridine orange, ethidium bromide, trypsin-EDTA, and crystal violet stain were purchased from Sigma Chemical Co. (St. Louis, MO, USA). Αlpha-hemolysin and b-tubulin antibodies were purchased from Abcam. Hematoxylin and eosin stains were purchased from Sigma (USA). All other chemicals and reagents were of the analytical grade level.