Bacterial strains utilized include Gram-negative: Salmonella typhimurium, (ATCC 13311).
Pseudomonas aeruginosa (ATCC 27853), Escherichia coli (ATCC 35210), and Gram-positive bacteria: Staphylococcus aureus (ATCC 6538) and Listeria monocytogenes (NCTC 7973) bacteria. Pathogens were provided from the Mycological Laboratory, Institute for Biological Research “Siniša Stankovic” Belgrade. Resistant strains used were MRSA IBRS MRSA 011, E. coli IBRS E003 and P. aeruginosa IBRS P001 obtained as described in Kartsev et al. [62]. The MIC/MBC were effectuated utilizing microdilution assay as previous described [63,64].
Sensitivity studies of E. coli strain were tested by the disc diffusion method on Mueller Hinton agar with the use of antibiogram discs (Bioanalyse) and tablets (Torlak, Serbia) for the following antibiotics: penicillin, amoxicillin, tetracycline, neomicin, gentamicin, colistin, ceftriaxon, sulfamethaxasole with trimetoprim, enrofloxacin and florfenicol. E. coli strain was resistant to all tested antibiotics with the exception of enrofloxacin, colistin and florfenicol [65]. It is described in detail in our previous paper [46].
Sensitivity studies of P. aeruginosa strain were tested by the disc diffusion method on Mueller Hinton agar with the use of antibiogram discs (Bioanalyse) and tablets (Torlak, Serbia) for the following antibiotics: penicillin, amoxicillin, tetracycline, neomicin, gentamicin, ceftriaxon, sulfamethaxasole with trimetoprim, enrofloxacin and florfenicol. P. aeruginosa strain was resistant to all tested antibiotics with the exception of enrofloxacin, and florfenicol [65,66].
This method was performed as described previously [48,67] with some modifications. The percentage of inhibition of biofilm formation was calculated by the following formula:
For the antifungal bioassays, six fungi were used: Aspergillus niger (ATCC 6275), Aspergillus fumigatus (human isolate), Aspergillus versicolor (ATCC 11730), Penicillium funiculosum (ATCC 36839), Trichoderma viride (IAM 5061), Penicillium verrucosum var. cyclopium (food isolate). The organisms were obtained from the Mycological Laboratory, Department of Plant Physiology, Institute for Biological Research ‘Siniša Stankovic’, Belgrade, Serbia. All experiments were performed in duplicate and repeated three times [68,69].
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