Insights into the flavor endowment of aroma-active compounds in cloves (Syzygium aromaticum) to stewed beef.

Clove (Syzygium aromaticum) is one of the most commonly used spices in stewed beef to enrich and improve its aroma during the stewing process. Gas chromatography ion mobility spectroscopy (GC-IMS), Q Exactive GC-Orbitrap-MS-O (QE-GC-MS/O), combined with sensory evaluation were employed to analyze the flavor endowment of aroma-active compounds in cloves to stewed beef. A total of 173 volatiles were identified in the clove powder (CP), stewed beef with clove (SBC), and stewed beef with salt (SBS), of which 21 volatiles were considered as aroma-active compounds. The concept of flavor endowment of aroma-active compounds in cloves was defined innovatively, and the endowment rate values (ERVs) of stewed beef were calculated. Nine aroma-active compounds in cloves were found to have a flavor endowment effect on stewed beef, while the terpenoids exhibited high ERVs. Despite the low ERV of eugenol, it still significantly impacted the aroma profile of SBC due to its high odor activity value (OAV) and flavor dilution (FD) factor. These volatiles offered mainly the clove, herbal, anise, and floral odor to stewed beef, which was also confirmed by sensory evaluation. These findings indicated that the terpenoids, phenolics and ethers in cloves had a significant influence on the overall aroma of stewed beef through the flavor endowment, which contributed to the precise use of cloves and improved the aroma of stewed beef.

Green Synthesis of Hexagonal-like ZnO Nanoparticles Modified with Phytochemicals of Clove (Syzygium aromaticum) and Thymus capitatus Extracts: Enhanced Antibacterial, Antifungal, and Antioxidant Activities.

The green synthesis of ZnO NPs is becoming increasingly valued for its cost-effectiveness and environmental benefits. This study successfully synthesized hexagonal ZnO NPs using a combination of clove (Syzygium aromaticum) and Thymus capitatus extracts. The use of both extracts significantly improved the antibacterial and antioxidant properties of the ZnO NPs. By optimizing synthesis conditions, including ZnCl2 and extract concentrations, hexagonal wurtzite ZnO NPs were produced at room temperature with only drying at 80 °C without high-temperature annealing. The synthesized ZnO NPs exhibited a hexagonal morphology with an average particle size of 160 nm and a crystallite size of 30 nm. Energy-dispersive X-ray spectroscopy (SEM-EDX) confirmed the elemental composition of the ZnO NPs, showing a high carbon content (63.9 wt.%), reflecting the presence of phytochemicals from the extracts coated the ZnO NPs surface. The UV-Vis spectrum revealed an absorption peak at 370 nm and a bandgap energy of 2.8 eV due to lattice defects caused by organic impurities. The ZnO NPs demonstrated exceptional antioxidant activity, with a DPPH radical scavenging rate of 95.2%. They also exhibited strong antibacterial activity against both Gram-positive and Gram-negative bacteria, with inhibition zones of 25 mm against Bacillus subtilis, 26 mm against Escherichia coli, 24 mm against Salmonella typhimurium, 22 mm against Klebsiella pneumoniae, 21 mm against Staphylococcus aureus, 20 mm against Staphylococcus hominis, and 18 mm against Bacillus subtilis at 200 ppm. Furthermore, significant antifungal activity was observed against Candida albicans, with an inhibition zone of 35 mm at the same concentration. These findings underscore the effectiveness of using combined plant extracts for producing ZnO NPs with controlled morphology and enhanced biological properties, highlighting their potential for various biomedical applications.

Cellulose synthesis from germinated tiger nut residue and its application in the production of a functional cookie.

In this study, cellulose was synthesized from the residue obtained after germinating tiger nuts for 0, 48, 72 or 96 h. The influence of the synthesized cellulose (0%, 2%, or 5%) on the quality of clove extract laden-cookies was evaluated. The optimum structure, morphology, and thermal properties of cellulose were obtained after geminating tiger nuts for 72 h. Adding cellulose to the dough stabilized the total phenolic, flavonoid, and protein contents and radical scavenging activity during the baking operation. The addition of 2% cellulose generally enhanced the hydration, pasting, and viscoelastic properties of the dough. However, 5% cellulose negatively affected the highlighted properties, culminating in poorer textural and sensory properties of the cookies produced therefrom. Germination could be effective in modifying the properties of cellulose from tiger nuts; thus, enhancing its application in the production of a functional cookie. Supplementary Information: The online version contains supplementary material available at 10.1007/s13197-024-05972-8.

Effects of clove bud oil on the physicochemical properties and oxidative stability of canola oil organogels structured by different beeswax contents.

The aim of this study was to investigate the utilization of clove bud oil as fat-soluble antioxidants for retarding lipid oxidation in organogels by structuring canola oil with beeswax at 5, 7.5, and 10% concentration under accelerated oxidation condition. Oil binding capacity and viscoelastic properties were increased with beeswax content, but were not nearly affected by the addition of clove bud oil. Organogel loaded with clove bud oil were found to be more effective in retarding lipid oxidation in high beeswax content systems, particularly evident in 10% beeswax samples. The addition of clove bud oil resulted in low levels of hyeoperxide and MDA, and protected against texture and color deterioration during the storage period. Additionally, the Pearson correlation between lipid oxidation indices and parameters of texture and color has been found to exhibit a limited association, with the exception of the a* and b* values, which show a strong correlation.

Clove essential oil and its major component, eugenol: a comparative study of their in vitro antioxidant and anticancer properties.

Syzygium aromaticum L. (clove) is a species native to subtropical countries. Its dried flower buds are rich in essential oils, which have shown insecticidal, anti-inflammatory and anaesthetic effects. This work was aimed to study the differences in antioxidant and anticancer activities between clove essential oil (CEO) and its major component, eugenol. The chemical composition of the CEO was determined by GC-MS. The physicochemical properties and antioxidant activity were determined in CEO and eugenol. Finally, anticancer activities were assayed against seven cell lines. Chemical analysis revealed that 80% of the CEO was eugenol. The density and IR were similar, and the colour was ΔE*>3. CEO had a lower phenolic content, but similar antioxidant activity to eugenol. The anticancer activity of the CEO was greater than that of eugenol in all the cell lines except for HeLa cells. These results suggest that secondary compounds in CEO enhance its antioxidant and -anticancer activities.

Development and Evaluation of Oromucosal Spray Formulation Containing Plant-Derived Compounds for the Treatment of Infectious and Inflammatory Diseases of the Oral Cavity.

According to data in the literature, natural products and essential oils are often used in dental practice. To develop a new oromucosal spray for the treatment of infectious and inflammatory diseases of the oral cavity, clove CO2 extract and essential oils of lavender and grapefruit were used as active pharmaceutical ingredients. Clove extract was obtained by the method of subcritical extraction from various raw materials, the choice of which was based on the yield of the CO2 extract and the study of its phytochemical and microbiological properties. Based on the results of microscopic and diffraction analyses, the rational time of ultrasonic exposure for the emulsion of active pharmaceutical ingredients was established. Mucoadhesive polymers were used as stabilizers of the two-phase system and prolongators. This article discusses the impact of the type and concentration of mucoadhesive polymers on the stability of the emulsion system; the viscous, textural, adhesive, and film characteristics of oromucosal spray; and the parameters determining sprayability.

Enhancing Deer Sous Vide Meat Shelf Life and Safety with Eugenia caryophyllus Essential Oil against Salmonella enterica.

Modern lifestyles have increased the focus on food stability and human health due to evolving industrial goals and scientific advancements. Pathogenic microorganisms significantly challenge food quality, with Salmonella enterica and other planktonic cells capable of forming biofilms that make them more resistant to broad-spectrum antibiotics. This research examined the chemical composition and antibacterial and antibiofilm properties of the essential oil from Eugenia caryophyllus (ECEO) derived from dried fruits. GC-MS analyses identified eugenol as the dominant component at 82.7%. Additionally, the study aimed to extend the shelf life of sous vide deer meat by applying a plant essential oil and inoculating it with S. enterica for seven days at 4 °C. The essential oil demonstrated strong antibacterial activity against S. enterica. The ECEO showed significant antibiofilm activity, as indicated by the MBIC crystal violet test results. Data from MALDI-TOF MS analysis revealed that the ECEO altered the protein profiles of bacteria on glass and stainless-steel surfaces. Furthermore, the ECEO was found to have a beneficial antibacterial effect on S. enterica. In vacuum-packed sous vide red deer meat samples, the anti-Salmonella activity of the ECEO was slightly higher than that of the control samples. These findings underscore the potential of the ECEO's antibacterial and antibiofilm properties in food preservation and extending the shelf life of meat.

Quality by design approach of apocynin loaded clove oil based nanostructured lipid carrier as a prophylactic regimen in hemorrhagic cystitis in vitro and in vivo comprehensive study.

Apocynin (APO) is a naturally occurring acetophenone with eminent anti-inflammatory and anti-oxidant peculiarities. It suffers from poor bioavailability due to low aqueous solubility. Herein, APO was loaded in a Clove oil (CO) based Nanostructured lipid carrier (NSLC) system using a simple method (ultrasonic emulsification) guided by a quality-by-design approach (23 full factorial design) to optimize the formulated NSLCs. The prepared NSLCs were evaluated regarding particle size (PS), polydispersity index (PDI), zeta potential (ZP), and entrapment efficiency (EE%). The optimal formula (F2) was extensively investigated through transmission electron microscope (TEM), Fourier transform infrared (FT-IR) spectroscopy, Differential scanning calorimetry (DSC), X-ray diffractometry (XRD), in vitro release, and stability studies. Cytotoxicity against human urinary bladder carcinoma (T24) cell line and in vivo activity studies in rats with induced cystitis were also assessed. The results disclosed that the optimal formula (F2) had PS of 214.8 ± 5.8 nm with EE% of 79.3 ± 0.9%. F2 also exhibited a strong cytotoxic effect toward the T24 cancer cells expressed by IC50 value of 5.8 ± 1.3 µg/mL. Pretreatment with the optimal formula (orally) hinted uroprotective effect against cyclophosphamide (CP)-induced hemorrhagic cystitis (HC) in rat models, emphasized by histopathological, immunohistochemical, and biochemical investigations. In consideration of the simple fabrication process, APO-loaded CO-based NSLCs can hold prospective potential in the prophylaxis of oncologic and urologic diseases.

Gelatin/polyvinyl alcohol films loaded with doubly stabilized clove essential oil chitosomes: Preparation, characterization, and application in packing marinated steaks.

In this study, a promising active food-packaging film of Gelatin/polyvinyl alcohol (GEL/PVA) integrated with doubly stabilized clove essential oil chitosome nanoparticles (CNP) was developed to maintain the freshness of marinated steaks. Results from the XRD and SEM experiments indicated excellent compatibility between the CNP and GEL/PVA matrix. Additionally, CNP was found to introduce more free hydroxyl groups, enhance the water retention and surface wettability of the CNP-GEL/PVA (C-G/P) film, and significantly reduce the swelling index from 963.78% to 495.11% (p < 0.05). Notably, the highest tensile strength and elongation at break (53.745 MPa and 46.536%, respectively) were achieved with the addition of 30% (v/v, based on the volume of gelatin) CNP; UVC was fully absorbed with 40% CNP; and films containing 60% CNP showed optimal inhibition of both Staphylococcus aureus and Escherichia coil, extending the shelf life of marinated steak from 3 to 7 days.

Identification of plant based potential antifungal compounds against BMK-1 protein of Bipolaris oryzae using molecular docking approach.

Rice brown spot is an important disease of rice worldwide that inflicts substantial yield losses. The antimicrobial potential of methanol, acetone and dimethyl sulfoxide (DMSO) extracts of different medicinal plants, viz., Syzygium aromaticum, Saussurea costus, Acorus calamus, Bergenia ciliate, Geranium pratense, Mentha longifolia, Inula racemosa, Podophyllum hexandrum, Heracleum candicans and Picrorhiza kurroa, against the brown spot pathogen Bipolaris oryzae in vitro was evaluated via mycelial growth inhibition and spore germination inhibition assays. Among the plant extracts tested, 100% mycelial inhibition was observed for the methanol extract of Syzygium aromaticum at all three concentrations (2000 ppm, 3000 ppm and 4000 ppm), followed by the methanol extract of Inula racemosa (90.33%) at 4000 ppm. A maximum conidial germination inhibition of 83.54% was exhibited by the Heracleum candicans leaf extract. Phytochemical profiling of Syzygium aromaticum and Inula racemosa through liquid chromatography and mass spectrometry (HR-LCMS) revealed the presence of several compounds, such as eugenol, ursolic acid, quercetin, chlorogenic acid, and noscapine. A molecular docking approach was used to identify key inhibitory molecules against B. oryzae. Among the compounds detected in S. aromaticum and Inula racemosa, ursolic acid and noscapine were found to have the greatest binding affinity for the Big Mitogen Activated Protein Kinase (BMK-1) enzyme present in B. oryzae. In conclusion, S. aromaticum and Inula racemosa are potent compounds that could serve as lead compounds for drug discovery in the future.

Synthesis and characterization of nano-biopolymer carriers loaded with clove (Syzygium aromaticum L.) extract as an anticancer agent: an in vitro study.

The objective of this work was to design a new drug nanoparticle (NP) composed of chitosan/ß-cyclodextrin/sodium tripolyphosphate/alginate (CS/ßCD/TPP/AL) loaded with a clove extract (CE) for potential anticancer effects. The extract was prepared by two extraction methods: hydroalcoholic maceration (MAC) with 80% MeOH and supercritical fluid (SCF). The MACCE and SCFCE CE NPs had particle sizes of 71 nm and 20 nm, respectively with irregular spherical shapes. The nanocarriers achieved entrapment efficiencies of over 90%. MACCE-NPs and SCFCE-NPs released 18.35% and 10.12% of the extract after 6 h, respectively. Cell viability decreased to 54%, 7%, and 12% in HeLa, U87, and KB cell lines, respectively, after a 48-hour treatment with SCFCE-NPs and 75%, 8%, and 17% after treatment with MACCE-NPs, significantly reduced compared to the control. It is concluded that NPs containing CE exhibit a higher degree of toxicity due to better penetration into cells.

Feasibility of Nanostructured Lipid Carrier Loaded with Alpha-Mangostin and Clove Oil for Canine Periodontal Therapy.

Nanostructured lipid carriers (NLC) represent the second generation of nanoparticles, offering numerous advantages over conventional delivery systems. These include improved stability, enhanced drug-loading capacity, and controlled release profiles, making them highly attractive candidates for a wide range of therapeutic applications. Their suitability for hydrophobic drugs like a traditional medicinal plant of Thailand as clove oil and alpha-mangostin. We investigated into nanostructured lipid carriers loaded with Alpha-Mangostin and clove oil (NLC-AMCO) into the physicochemical and biological characteristics to identify the formulation with the highest efficacy for treatment. The particle size, charge, polydispersity index, and other characterizations were recorded. The realtime ex vivo penetration was explored using canine gingival tissue. Drug sustained release was assessed by HPLC. Moreover, the antibacterial properties were tested by conventional methods. The NLC-AMCO can be stored at up to 40 °C for 60 days without any alterations in particle characteristics. Gingival tissue penetration and sustained drug release were superior compared to unencapsulated counterparts. It exhibited greater effectiveness in inhibiting bacterial growth than the antibiotics tested, particularly against bacteria from the oral cavities of dogs. Therefore, this alternative treatment approach offers cost-effectiveness and ease of administration for pet owners and reduces discomfort for the animals during restraint.

Preparation and functional characterization of pullulan-sodium alginate composite film enhanced with ultrasound-assisted clove essential oil Nanoemulsions for effective preservation of cherries and mushrooms.

Clove essential oil (CEO) exhibited potent antibacterial efficacy and are obtained from Eugenia caryophyllata tree flower buds. Herein, CEO nanoemulsions were prepared using various concentrations of casein protein treated with ultrasound for different time interval. The study demonstrated that CEO nanoemulsions with 5% casein protein subjected to ultrasound for 10 min displayed the most minimal particle size. The pullulan­sodium alginate film incorporated with nanoemulsions treated with ultrasound exhibited enhanced physico-mechanical characteristics. Based on the structural analysis, the application of ultrasonic treatment improved intermolecular compatibility and organized molecular structure by strengthening hydrogen bonds. Furthermore, the composite film displayed remarkable efficacy against E. coli and S. aureus as well as longer retention of essential oils. The use of the developed films to protect cherry fruits and mushrooms produced promising results, emphasizing their potential in food packaging applications.

Development of optimized functional clove fortified probiotic yoghurt.

We focused on assessing the antimicrobial effects of functional yoghurts supplemented with clove and probiotics. The formulation of aqueous clove extract (ACE) incorporated with probiotic yoghurt (Streptococcus thermophilus, Lactobacillus bulgaricus, and Lactococcus lactis) was optimised in terms of aqueous clove extract concentrations (2.5-7.5% v/v), fermentation temperature (32-42 °C), and total culture concentration (1.5-4.5% v/v). pH, titratable acidity, syneresis, water holding capacity, viscosity, springiness, color difference, lactic acid bacteria viability, and the antibacterial property of 17 runs were determined as responses using Box-Behnken design. The results indicate that elevated ACE concentration leads to a significant (p < 0.05) increase in titratable acidity, antibacterial effectiveness (against K. pneumoniae and P. aeruginosa), springiness, and color profile. Conversely, an elevated fermentation temperature significantly (p < 0.05) reduces pH and L. bulgaricus viability (log CFU/mL). Additionally, there is a significant (p < 0.05) decline in S. thermophilus and L. lactis viability (log CFU/mL) as well as springiness with an increased culture concentration. The optimal conditions identified are 7.5% (v/v) ACE concentration, a fermentation temperature of 36.6 °C (37 °C), and a total culture concentration of 4.5% (v/v), resulting in a 79% desirability score. The spectra of components were mainly obtained at wavelength of 3258 cm-1, 1636 cm-1 and 1075 cm-1 in Fourier transform infrared (FTIR) spectroscopy of optimized functional yoghurt. Where, 3258 cm-1 corresponds to the stretching vibration of O-H (hydroxyl) groups, 1636 cm-1 corresponds to the C=O (carbonyl) stretching vibration, and 1075 cm-1 corresponds to the C-O (ether or alcohol) stretching vibration.

Effects of clove oil concentrations on blood chemistry and stress-related gene expression in Siamese fighting fish (Betta splendens) during transportation.

Siamese fishing fish (Betta splendens) or betta are usually subjected to a special method of transportation for global trade, where they are individually conveyed in plastic bags containing just enough water to cover their bodies. This study aimed to investigate the effects of transportation on their stress response by measuring hematological values, stress hormone levels, glucose levels, and stress-related gene expression. Betta fish (average body weight 1.91 ± 0.42 g; n = 30) were exposed to simulated transport in a water volume of 40 mL for 12, 24, and 48 h. Baseline levels (pre-transport) were measured prior to the experiment. The control group was transported using water without adding clove oil. Two treatment groups were transported using water with the addition of 1 and 3 mg/L concentrations of clove oil, respectively. The results revealed that transportation can be a factor that affects water quality. The pH and dissolved oxygen levels were significantly lower than baseline, while nitrite and total ammonia concentrations significantly increased. Correlating to the stress responses, significantly increasing total red blood cell counts, plasma cortisol levels, and up-regulating the expression of stress-related genes, including HSP70, GR, MR, and HIF-1α. The addition of 1 mg/L clove oil was found to reduce stress during the transport simulation, as evidenced by a reduction in these stress parameters. Conversely, increasing the concentration of clove oil to 3 mg/L significantly increased plasma cortisol after 12 h of simulated transport, and up-regulated GR, MR, and HIF-1α expression. This study revealed that the transport process can stimulates stress in betta fish but adding a concentration of 1 mg/L clove oil to the transport water could mitigate this stress response and promote animal welfare during their transportation.

Effect of eugenol on cytochrome P450 1A2, 2C9, 2D6, and 3A4 activity in human liver microsomes.

This study aimed to provide an understanding of the influence of eugenol on CYP1A2, 2C9, 2D6, and 3A4 in human liver microsomes (HLM). Specific substrate for CYP1A2, 2C9, 2D6, and 3A4 were incubated in HLM with or without eugenol. The formation of their respective metabolites was assessed with HPLC analytical methods. Eugenol at 1, 10 and 100 µM levels inhibited the activity of CYP1A2 and CYP2C9 by 23.38 %, 23.57 %, 39.80 % and 62.82 %, 63.27 %, 67.70 % respectively. While, CYP2D6 and CYP3A4 activity was decreased by 40.70 %, 45.88 %, 62.68 % and 37.41 %, 42.58 % and 67.86 % at 1, 10 and 100 µM eugenol level respectively. The IC50 value of eugenol for CYP2D6 and CYP3A4 was calculated as 11.09 ± 3.49 µM and 13.48 ± 3.86 µM respectively. Potential herb-drug interactions was noted when eugenol is administered simultaneously with medications metabolized by these enzymes, most notably CYP2C9, CYP2D6 and CYP3A4.

Preparation of Fresh-Keeping Paper Using Clove Essential Oil through Pickering Emulsion and Maintaining the Quality of Postharvest Cherry Tomatoes.

This study focused on developing a Pickering emulsion fresh-keeping paper that contained clove essential oil (CEO). Cherry tomatoes served as the test material for assessing the preservative efficacy of fresh-keeping paper. The results showed that Pickering emulsion had strong stability. Additionally, the fresh-keeping paper had a good antioxidant activity and sustained-release effect on CEO. In terms of the preservation effect, 0.75 wt% CEO Pickering emulsion paper reduced the decay incidence and weight loss of cherry tomatoes during 12-day storage. Fresh-keeping paper could also play a positive role in protecting the sensory index and color difference of tomatoes. It slowed the decline rate of soluble solid concentration (SSC) and titrable acid (TA). The vitamin C (Vc) and hardness of preserved tomatoes using fresh-keeping paper were maintained at a high level. The paper also inhibited the growth of microorganisms significantly. Therefore, 0.75 wt% CEO Pickering emulsion fresh-keeping paper displayed considerable potential for application in the preservation of postharvest fruits and vegetables. It is a novel fruit and vegetable preservation material worthy of development.

Insights from Syzygium aromaticum Essential Oil: Encapsulation, Characterization, and Antioxidant Activity.

Alginate encapsulates loaded with clove essential oil (CEO) were prepared by ionic gelation, with subsequent freeze-drying. The objective of the present work was to develop a product with the ability to protect CEO against its easy volatility and oxidation. The following techniques were used to characterize the formulations: eugenol release, degree of swelling, GC/MS, TGA/DSC, and SEM. The alginate solution (1.0%) containing different concentrations of CEO (LF1: 1.0%; LF2: 0.5%; LF3: 0.1%) was dropped into a 3.0% CaCl2 solution. After lyophilization, the encapsulated samples were wrinkled and rigid, with high encapsulation power (LF3: 76.9% ± 0.5). Three chemical components were identified: eugenol (the major one), caryophyllene, and humulene. The antioxidant power (LF1: DPPH IC50 18.1 µg mL-1) was consistent with the phenol content (LF1: 172.2 mg GAE g-1). The encapsulated ones were thermally stable, as shown by analysis of FTIR peaks, eugenol molecular structure was kept unaltered. The degree of swelling was 19.2% (PBS). The release of eugenol (92.5%) in the PBS solution was faster than in the acidic medium. It was concluded that the low-cost technology used allows the maintenance of the content and characteristics of CEO in the three concentrations tested, offering a basis for further research with essential oil encapsulates.

Flaxseed Mucilage/Hydroxypropyl Methylcellulose and Sodium Alginate/Polyvinyl Alcohol Composite Bilayer Film as a Promising Drug Carrier for Periodontal Treatment.

Objectives: The present study focused on the formulation of mucoadhesive bilayer composite films for the treatment of periodontitis and evaluation of their physicochemical properties. Materials and Methods: The solvent casting technique was used to prepare films. The primary layer (D) was prepared with flaxseed and hydroxypropyl methylcellulose composite to sustain the release of doxycycline hyclate. The second layer (S) comprised sodium alginate and polyvinyl alcohol composite for faster release of clove oil. Both layers were combined to generate the bilayer film (B). All formulations were characterized further to obtain an optimized formulation. Results: Attenuated total reflection-Fourier transform infrared radiation results showed intactness of drug and clove oil in the presence of excipients. The pH of the films was compatible with the periodontal cavity and the thickness was suitable for inserting into the cavity. The immediate release layer showed faster disintegration and swelling. The content of clove oil was above 80%. The rate of swelling of the primary layer was slow and drug content complied with the United States Pharmacopoeia. Scanning electron microscope analysis revealed intact, non-porous and smooth films. Films exhibited better mechanical strength and bioadhesiveness. Clove oil was released from the immediate release layer within 10 min, and doxycycline hyclate release was retarded to a minimum of up to 8 h in the primary layer as well as the bilayer. Formulation also had a significant effect on both Escherichia coli and Staphylococcus aureus. Conclusion: In the current study, bilayers were successfully prepared and characterized. The optimized formulation can be effectively used for the treatment of periodontitis.

Safety and efficacy of a feed additive consisting of a tincture derived from the flowers of Syzygium aromaticum (L.) Merr. & L.M. Perry (clove tincture) for all animal species (FEFANA asbl).

Following a request from the European Commission, EFSA was asked to deliver a scientific opinion on the safety and efficacy of a tincture from the dried flower bud of Syzygium aromaticum (L.) Merr. & L.M. Perry (clove tincture) when used as a sensory additive in feed and water for drinking for all animal species. The product is a ■■■■■) solution, with a dry matter content of ~ 1.66%. The product contains on average 0.511% phenolic acids (of which 0.0344% were flavonoids), 0.039% eugenol, 0.00019% methyleugenol and 0.00008% estragole. The Panel on Additives and Products or Substances used in Animal Feed (FEEDAP) concluded that the use of clove tincture is very unlikely to be of safety concern for the target species up to the maximum proposed use level of 50 mg clove tincture/kg complete feed for all animal species, except for horses, for which the proposed use level is 200 mg/kg complete feed. The FEEDAP Panel considers that the use in water for drinking alone or in combination with use in feed should not exceed the daily amount that is considered very unlikely to be of safety concern when consumed via feed alone. No safety concern would arise for the consumer and the environment from the use of clove tincture up to the maximum proposed use levels in feed. The additive under assessment should be considered as irritant to skin and eyes, and as a dermal and respiratory sensitiser. When handling the additive, exposure of unprotected users to methyleugenol and estragole may occur. Therefore, to reduce the risk, the exposure of the users should be minimised. Since the flower buds of S. aromaticum and their preparations were recognised to flavour food and their function in feed would be essentially the same, no demonstration of efficacy was considered necessary.