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Kefiran is a heteropolysaccharide that is considered a postbiotic and is obtained by kefir grains fermented in cow’s milk, while little is known about the donkey milk (DM) variety. Postbiotics are recognised as having important human health benefits that are very similar to probiotics but without the negative effects associated with their ingestion. Donkey is a monogastric animal, as are humans, and when used as an alternative food for infants who suffer from cow milk protein allergies, DM could therefore display more biocompatibility. In this study, the DM kefiran was extracted by ultrasound from kefir grains cultured in donkey milk and fully characterized for its structural and physicochemical properties by Fourier-transform infrared spectroscopy (FT-IR), High-Performance Liquid Chromatography- Refractive Index (HPLC-RI), Scanning electron microscope (SEM), Differential Scanning Calorimeters (DSC) and rheological analyses. In addition, tests were conducted on keratinocytes cell lines and human red blood cells to assess the nontoxicity and haemolysis degree of the polymer. The extraction yield of the DM kefiran was 6.5 ± 0.15%. The FT-IR analysis confirmed the structure of the polysaccharide by showing that the stretching of the C-O-C and C-O bonds in the ring, which formed two bands at 1157 and 1071 cm−1, respectively, and the anomeric band at 896 cm−1 indicates the β configuration and vibrational modes of glucose and galactose. Results were confirmed by HPLC-RI analysis indicating that the ratio glucose/galactose was 1:0.87. Furthermore, the SEM analysis showed a porous and homogeneous structure. The rheological analysis confirmed the pseudoplastic nature of the polymer, while the DSC analysis highlighted excellent thermal resistance (324 °C). Finally, DM kefiran was revealed to have biologically acceptable toxicity, showing a haemolytic activity of less than 2% when using fresh human red blood cells and showing no cytotoxicity on human keratinocytes. Therefore, kefiran obtained by DM shows an excellent biocompatibility, establishing it as a promising polymer for bioengineering human tissue for regenerative applications.

Active packaging films based on kefiran-gelatin were developed and characterized using Zhumeria majdae essential oil nanoemulsion (ZMEO-NE) at concentrations of 0 (control), 1, 2 and 3 %. The main compounds of the essential oil (EO) of Zhumeria majdae (ZM) plant were linalool (61.44 %) and camphor (20.67 %). Adding the ZMEO-NE to the films decreased permeability to water vapor (from 7.82 × 10 -7 to 4.09 × 10 -7 g·m/m 2 ·Pa·h), ultimate tensile strength (from 38.44 to 33.48 MPa), percentage of light transmission, and increased thickness (from 0.085 to 0.121 mm), opacity (from 2.11 to 2.79), and elongation at break (from 19.97 to 34.73 %), and changed color parameters. The establishment of hydrogen bonds between the ZMEO-NE and polymer network was confirmed. The ZMEO-NE decreased the storage modulus and glass transition temperature. Distinct variations in the films' surface morphology and a reduction in the crystalline structure were observed due to the presence of the ZMEO-NE. Elevating the concentration of the ZMEO-NE increased antioxidant capabilities of the films. The films incorporating the ZMEO-NE exhibited notable antibacterial efficacy against Staphylococcus aureus (reductions ≥4 log CFU/cm 2 ) and Escherichia coli (reductions ≥2 log CFU/cm 2 ) bacteria. The films also demonstrated suitable antifungal properties during storage of sponge cakes for 16 days., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2024 Elsevier B.V. All rights reserved.)