Campylobacter is the leading cause of bacterial foodborne gastroenteritis worldwide. Infections by Campylobacter in humans are generally caused by consuming contaminated foods of animal origin, with poultry, especially chicken, being the main reservoir. The high prevalence of Campylobacter in chicken carcasses and the growing resistance to the most widely used antibiotics has driven EFSA to propose a regulation (2017/1495) containing new microbiological criteria to regulate the presence of Campylobacter in broiler carcasses. In this context, there has been an increase in the number of research aimed at the search for new tools to reduce Campylobacter incidence in chicken meat. The objective of the present work was to evaluate the antibacterial activity of two olive leaf extracts (A y B) against eleven Campylobacter spp. strains (C. jejuni y C. coli) isolated from chicken food chain. Results showed that all Campylobacter strains had resistance to at least one of the eight antibiotics evaluated, and 46% of them were antibiotic multi-resistant. HPLC analysis showed that hydroxytyrosol and oleuropein were the major phenolic compounds in extracts A and B, respectively. Extract A showed a significant antibacterial activity against all Campylobacter strains tested in the present work. The use of a pure standard of hydroxytyrosol confirmed the contribution of this compound to the antibacterial effect of extract A. These results suggest that olive leaves could be used as a source of bioactive compounds to obtain extracts with antibacterial activity against Campylobacter spp potentially applicable to reduce the presence of Campylobacter in chicken carcasses.

Consumption of omega-3 long chain polyunsatured fatty acids (PUFAs) abundant in oily fish, krill, shrimp, and algae is critical for the physical and mental health of adults and children [1]. Called by Winkler the most hidden of all the hidden hungers [2], the populations of most world’s countries share an insufficient daily intake of both eicosapentaenoic acid (EPA) and docosahexenoic acid (DHA). This diet deficiency led national health authorities to recommend daily intakes of both PUFAs either by increasing consumption of fish and crustaceans or by intake of omega-3 dietary supplements. Besides a minor share of the industry using omega-3 lipids extracted from algae, the large and growing omega-3 food supplement industry uses refined fish or krill oil as raw material. Increasing demand of fish oil adds to the overfishing pressure threatening many species, including anchovies, menhaden and krill [3]. The conventional fish oil extraction involves a multi-step, energy-intensive process starting on board of the shipping vessel where once caught anchovies are cooked and pressed giving an oil in water suspension. After reaching the industrial site, the oily mixture undergoes centrifugation and subsequent chemical refinement eventually affording EPA and DHA in ethyl ester form [4]. Besides contributing to overfishing, the process removes from the refined oil important antioxidant compounds such as carotenoids and biophenols that protect chemically labile PUFAs from oxidation and autooxidation [5]. We recently introduced a circular and green method for the production of fish oil rich in omega-3 from the leftovers of anchovy fillets based on solid-liquid extraction using d-limonene as biosolvent [6]. The resulting fish oil contains both EPA and DHA in their natural (triglyceride) form, along with significant levels of vitamin D₃ [7]. The use of fishery byproducts, as raw materials for the production of fish oil omega-3 extracts is highly desirable. Renewably obtained from waste orange peel, the biosolvent limonene is nearly entirely recovered after the extraction whereas its antimicrobial, antifungal, and antioxidant properties protect the PUFAs during the extraction ensuring high recovery rate also of the natural antioxidant zeaxanthin abundant in anchovies [8]. The method closes the materials cycle and establishes a circular economy process to obtain high quality fish oil from bio-based waste available worldwide in several million t/year amount. The extraction of omega-3 lipids from anchovy discards using d-limonene as only solvent, indeed, is economically and technically feasible on large scale [8].

References

[1] F. Shahidi, P. Ambigaipalan, Omega-3 polyunsaturated fatty acids and their health benefits, Annu. Rev. Food Sci. Technol. 2018, 9, 345−381.

[2] J.T. Winkler, The most hidden of all the hidden hungers: the global deficiency in DHA and EPA and what to do about it, World Rev. Nutr. Diet. 2018, 118, 123−130.

[3] P. Greenberg, The Omega Principle, Penguin Press, New York: 2018.

[4] R. Ciriminna, F. Meneguzzo, R. Delisi, M. Pagliaro, Enhancing and improving the extraction of omega-3 from fish oil, Sustainable Chem. Pharm. 2017, 5, 54−59.

[5] M. Pagliaro, D.M. Pizzone, A. Scurria, C. Lino, E. Paone, F. Mauriello, R. Ciriminna, Sustainably sourced olive polyphenols and omega-3 marine lipids: a synergy fostering public health, ACS Food Sci. Technol. 2021, 1, 139-145.

[6] R. Ciriminna, A. Scurria, G. Avellone, M. Pagliaro, A circular economy approach to fish oil extraction, ChemistrySelect 2019, 4, 5106-5109.

[7] A. Scurria, C. Lino, R. Pitonzo, M. Pagliaro, G. Avellone, R. Ciriminna, Vitamin D₃ in fish oil extracted with limonene from anchovy leftovers, Chem. Data Collect. 2020, 25, 100311.

[8] R. Ciriminna, A. Scurria, A. S. Fabiano-Tixier, C. Lino, G. Avellone, F. Chemat, M. Pagliaro, Omega-3 extraction from anchovy fillet leftovers with limonene: chemical, economic and technical Aspects, ACS Omega 2019, 4, 15359-15363.

The INFOGEST digestion method was adapted for the evaluation of sterol bioaccessibility in a plant sterol (PS)-enriched beverage [1]. Recent modifications of the method such as the addition of gastric lipase and cholesterol esterase, showed that the presence of sterols in the digestion reagents has an important role on sterol bioaccessibility [2]. However, the specific sterol contribution from these extracts is unknown. Therefore, in the present study, the sterol content of the extracts used in the INFOGEST 2.0 digestion method (rabbit gastric extract – RGE –, porcine pancreatin and bovine bile) is determined by GC-FID. Results show that cholesterol content of the extracts is as follows: bovine bile > porcine pancreatin > RGE (4.07±0.18, 1.41±0.07 and 0.57±0.05 mg/g extract, respectively). Considering the amount of each extract added to the digestion, the greatest cholesterol contributor is porcine pancreatin followed by bovine bile and RGE (1.72±0.08, 1.00±0.04, and 0.046±0.004 mg, respectively). However, cholesterol in bile is found in pre-formed micelles that increase its solubility, and, therefore, it would compete for sterol micellarization against sterol provided by the digested foods. This fact has been observed in previous studies in which the cholesterol content of the blanks of digestion is inversely correlated with PS bioaccessibility [2]. On the other hand, bovine bile contains stigmasterol, β-sitosterol and sitostanol (0.14±0.01, 0.93±0.07 and 0.18±0.01 mg/g extract, respectively), while porcine pancreatin contains campesterol, stigmasterol, β-sitosterol and sitostanol (0.103±0.004, 0.25±0.02, 2.17±0.42 and 0.32±0.01 mg/g extract, respectively). Nevertheless, these PS present in the extracts appear in negligible amounts in the digestion blank, reflecting their low solubility vs. cholesterol. In conclusion, in order to optimize the INFOGEST 2.0 gastrointestinal method for the evaluation of sterol bioaccessibility, the characterization of the sterol content in the digestion reagents provides valuable information since it may condition their solubility.

References: [1]. Blanco-Morales et al. Food Funct, 2018, 9, 2080-2089; [2]. López-García et al. 2021, Book of abstracts “Virtual International Conference on Food Digestion”, pp 44 (https://www.cost-infogest.eu/content/download/4051/35805/file/V-ICFD%20Book%20of%20Abstracts.pdf)

Acknowledgements: Authors thank the financial support from project PID2019-104167RB-I00/AEI/10.13039/501100011033 (Ministry of Science and Innovation, Spain). Mussa Makran holds an FPU19/00156 grant from the Ministry of Universities (Spain). Gabriel López-García holds a junior doctor researcher contract within the aforementioned project (CPI-17-025).

Background: Medicinal plants continue to be an interesting source of natural products for treating various health conditions. It is estimated that more than 150,000 plant species have been studied, many of which contain valuable therapeutic agents, and the applications of novel compounds from plants for pharmaceutical purposes have been gradually increasing in recent years.
Essential oils are volatile compounds with an oily fragrance obtained from the different plant parts, flowers, leaves, stems, bark, and roots. These volatile oils showed the different kinds of biological activities, including antibacterial, antioxidant, antiviral, insecticidal, etc. The importance of essential oils is increasing because they are mostly used in the beverage and food industries, cosmetics and fragrance industries to make valuable perfumes, and with many biological activities.
Medicinal and aromatic plants are extensively used as natural organic compounds and as medicines. Also, essential oils of aromatic plants have been used to treat various sorts of infectious diseases. In recent years, numerous efforts are made to explore essential oils used to treat various diseases.
Vitex rotundifolia L. is a deciduous shrub belonging to the Verbenaceae. Its fruits have been used in traditional medicine to treat colds, headaches, chronic otitis media, neuralgia, and various diseases. The extractives of Vitex rotundifolia L contain various chemical components, such as flavonoids, diterpenes, terpenoids, and lignans. In addition, it has various bioactivities such as antioxidant, anti-cancer, anti-inflammatory, anti-allergic, and soothing effects.
Objective: The purpose of this study was to determine the potential as an inhibitor of mucus production through evaluating the anti-asthmatic effect of essential oil from V. rotundifolia fruit.
Methods: The essential oil was extracted from V. rotundifolia fruit by hydrodistillation method. Chemical compositions of the essential oil were identified by GC-MS analysis. GC-MS was equipped with a VF-5MS column.
The anti-asthmatic activity of essential oil was examined using the LPS-stimulated NCI-H292 cells. NCI-H292 cells were cultured in a 37℃, 5% CO₂ incubator, followed by treatment with various concentrations (10^-3%~10^-7%) of essential oil from V. rotundifolia fruit. After 24 h incubation, CCK-8 solution was added to each well, and the 96-well plate was incubated at 37℃ for 1 h. The percentage of cells showing was determined relative to the control group. Relative gene expression changes of MUC4, MUC5AC, and MUC5B were investigated in the LPS-stimulated NCI-H292 cells treated with essential oil.
Results: As a result of GC/MS analysis, α-pinene (38.07%), 1,8-cineol (12.73%), 5α,9α,10β-kaur-15-ene (7.53%), β-pinene (6.20%), and α-terpinyl acetate (5.22%) were identified as major components of the essential oil. The essential oil exhibited toxicity in a dose-dependent manner. The cell viability was higher than 80% at oil concentrations of 10-7%~10-5%, and almost no cytotoxicity was observed at a concentration lower than 10-5% in NCI-H292. When LPS-stimulated NCI-H292 cells were treated with essential oil (10^-7%), the relative expression levels of the mucus genes MUC4, MUC5AC, and MUC5B were significantly reduced compared to the LPS-treated group (NC).
According to the results of the relative expression level of mucus genes, the expression level of the MUC4, membranous mucin, was decreased by about 42% compared to the negative control. The relative expression of MUC5AC, the most important secretory mucus gene among the mucus genes, was reduced by about 73.6% compared to the negative control. In MUC5B, another secretory mucinous, the essential oil inhibited the gene expression of MUC5B effectively. The MUC5B expression was decreased by about 69.3% in the LPS-stimulated NCI-H292 cells. Budesonide, the positive control, was reduced the relative expression of mucin genes MUC4, MUC5AC, and MUC5B by 7.4%, 18.6%, and 86.6%, respectively. The anti-asthmatic activity of essential oil was very good compared with budesonide.
Conclusion: The anti-asthmatic effect of essential oil from V. rotundifolia fruit was confirmed. The anti-asthmatic activity of essential oil was superior to budesonide, an anti-asthmatic treatment. These results are suggested that that the essential oil of V. rotundifolia fruit can reduce asthma response by suppressing the expression of the mucus gene.

People are using plants with health-boosting properties, such as oregano species, in their daily diet. Nowadays these plants, characterized as potential functional foods, are being extensively scientifically analyzed. Bearing that in mind and the fact that neurodegenerative diseases such as Alzheimer’s and Parkinson’s are affecting millions of people worldwide, this study was defined to assess the antineurodegenerative, antineuroinflammatory, and neuroprotective potential of Origanum majorana (marjoram) and O. vulgare (oregano) from Serbia. The plant material was obtained from the Institute for Medicinal Plant Research “Dr. Josif Pančić”, Serbia, and the ethanolic extracts were prepared using classic maceration protocol. The acetylcholinesterase (AChE) and tyrosinase (TYR) inhibition were used to examine the antineurodegenerative activity. Microglial (BV2) cells stimulated with LPS were used to evaluate the antineuroinflammatory activity in MTT, NBT, and Griess assays, while neuronal (SH-SY5Y) cells were used to determine the neuroprotective activity of the extracts in MTT and Griess assays. The results suggest that oregano extracts are slightly better at inhibiting AChE and TYR (68% vs. 61% at 0.1 mg/mL) than marjoram extracts (53% vs. 59% at 0.1 mg/mL), making them more active than the positive controls (galantamine 41% vs. kojic acid 25% at 0.1 mg/mL). Furthermore, these extracts normalized LPS-stimulated BV2 cells' viability while also reducing their production of inflammatory mediators, reactive oxygen species, and nitric oxide, to the level of untreated cells. Additionally, supernatants of LPS-stimulated BV2 cells that were previously treated with these extracts normalized the viability of neurons compared to the control neurons (treated with supernatants of LPS-stimulated BV2 that were not previously treated with the extracts). Both extracts exhibited noticeable antineurodegenerative antineuroinflammatory and neuroprotective activities representing powerful sources of phytochemicals with promising overall neuroprotective activity, which could be further examined for potential dietary supplement manufacturing.