Research progress on plant stress-associated protein (SAP) family: Master regulators to deal with environmental stresses.

Every year, unfavorable environmental factors significantly affect crop productivity and threaten food security. Plants are sessile; they cannot move to escape unfavorable environmental conditions, and therefore, they activate a variety of defense pathways. Among them are processes regulated by stress-associated proteins (SAPs). SAPs have a specific zinc finger domain (A20) at the N-terminus and either AN1 or C2H2 at the C-terminus. SAP proteins are involved in many biological processes and in response to various abiotic or biotic constraints. Most SAPs play a role in conferring transgenic stress resistance and are stress-inducible. The emerging field of SAPs in abiotic or biotic stress response regulation has attracted the attention of researchers. Although SAPs interact with various proteins to perform their functions, the exact mechanisms of these interactions remain incompletely understood. This review aims to provide a comprehensive understanding of SAPs, covering their diversity, structure, expression, and subcellular localization. SAPs play a pivotal role in enabling crosstalk between abiotic and biotic stress signaling pathways, making them essential for developing stress-tolerant crops without yield penalties. Collectively, understanding the complex regulation of SAPs in stress responses can contribute to enhancing tolerance against various environmental stresses through several techniques such as transgenesis, classical breeding, or gene editing.

Enhancing the Shelf Life of Sous-Vide Red Deer Meat with Piper nigrum Essential Oil: A Study on Antimicrobial Efficacy against Listeria monocytogenes.

Using sous-vide technology in combination with essential oils offers the potential to extend the preservation of food items while preserving their original quality. This method aligns with the growing consumer demand for safer and healthier food production practices. This study aimed to assess the suitability of minimal processing of game meat and the effectiveness of vacuum packaging in combination with Piper nigrum essential oil (PNEO) treatment to preserve red deer meat samples inoculated with Listeria monocytogenes. Microbial analyses, including total viable count (TVC) for 48 h at 30 °C, coliform bacteria (CB) for 24 h at 37 °C, and L. monocytogenes count for 24 h at 37 °C, were conducted. The cooking temperature of the sous-vide was from 50 to 65 °C and the cooking time from 5 to 20 min. Additionally, the study monitored the representation of microorganism species identified through mass spectrometry. The microbiological quality of red deer meat processed using the sous-vide method was monitored over 14 days of storage at 4 °C. The results indicated that the TVC, CB, and L. monocytogenes counts decreased with the temperature and processing time of the sous-vide method. The lowest counts of individual microorganism groups were observed in samples treated with 1% PNEO. The analysis revealed that PNEO, in combination with the sous-vide method, effectively reduced L. monocytogenes counts and extended the shelf life of red deer meat. Kocuria salsicia, Pseudomonas taetrolens, and Pseudomonas fragi were the most frequently isolated microorganism species during the 14-day period of red deer meat storage prepared using the sous-vide method.

Physico-Chemical Properties and Chemical Analysis of Wildflower Honey Before and After the Addition of Spirulina (Arthrospira platensis).

In this study, in order to verify the effects due to the addition of spirulina (Arthrospira platensis) in a food product, a wildflower honey was analyzed in terms of chemical composition, physicochemical properties and antioxidant activity before and after the addition of the spirulina. HS-SPME/GC-MS and HPLC/UV were applied to carry out the chemical analyses. The obtained results demonstrated that the volatile profile and also the sugar content were significantly influenced by the addition of spirulina, showing significant qualitative and quantitative differences compared to honey without spirulina. The increase in HMF in honey added with spirulina was significant, demonstrating that its presence could accelerate the Maillard reaction. Electrical conductivity measured by using a conductometer was also increased while the moisture content was reduced in honey enriched with spirulina. Instead, the pH value was similar between the two samples. On the other hand, honey fortification with spirulina determined a significant increase of 12.5% in the total phenolic content (TPC), and a 56.25% increase in protein content. Further, the total antioxidant capacity (TAC) was also evaluated and a significant increase was determined as a result of the addition of spirulina. In conclusion, honey enriched with A. platensis was found to be characterized by a high pool of bioactive metabolites as well as significant changes in almost all the measurements performed.

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.

LVI and DI-SPME Combined with GC/MS and GC/MS for Volatile Chemical Profile Investigation and Cytotoxic Power Evaluation of Essential Oil and Hydrolate from Cannabis sativa L. cv. Carmagnola.

Cannabis sativa L. is a plant that has been cultivated since ancient times thanks to its various uses. Even its extraction products, such as essential oil and hydrolate, having a varied chemical composition and rich in bioactive components, find wide use in different sectors, gathering ever-increasing interest over time. In this work, the essential oil of Cannabis sativa L. cv. Carmagnola was characterized by using Gas Chromatography/Mass Spectrometry (GC/MS) and, for the first time, the chemical profile of the hydrolate was also described through different analytical techniques such as Large-Volume Injection Gas Chromatography/Mass Spectrometry (LVI-GC/MS) and Direct Immersion-Solid Phase Microextraction-Gas Chromatography/Mass spectrometry (DI-SPME-GC/MS), in order to provide a more complete compositional profile. The results of the analyses conducted on the hydrolate highlighted a high content of α-terpineol; on the other side, in the essential oil, a prevalence of monoterpenes, with α-pinene and limonene as the characterizing components, was detected. Both matrices were also investigated to evaluate their cytotoxic activity by using a panel of cancer cell lines derived from different histotypes such as melanoma (A375, LOX IMVI), non-small cell lung cancer (H1299, A549), colon (HT29) and pancreatic (L3.6) cancer cell lines. The obtained data demonstrated that essential oil was more effective than hydrolate in terms of reduction in cell viability.

Anti-inflammatory and antiarthritic properties of Marrubium vulgare aqueous extract in an animal model.

This study aimed to investigate the potential anti-inflammatory properties of aqueous extract of Marrubium vulgare (AEMV) using various animal models. Several inflammatory models including xylene-induced ear edoema, carrageenan-induced paw edoema, and Freund's adjuvant-induced arthritis were employed to evaluate the anti-inflammatory effects of AEMV. LC-MS/MS of AEMV revealed that the major component was Marrubiin, a diterpenoid lactone. AEMV demonstrated significant anti-inflammatory effects in all animal models tested. It effectively reduced ear and paw edoema induced by xylene and carrageenan, respectively. Furthermore, AEMV attenuated arthritis symptoms and hyperalgesia in rats with Freund's adjuvant-induced arthritis. Biochemical analyzes revealed normalisation of inflammatory markers, including C-reactive protein (CRP) levels, in treated animals. The findings suggest that AEMV possesses promising anti-inflammatory properties, supporting its potential therapeutic application in inflammatory conditions such as arthritis. Further investigations are needed to clarify the underlying mechanisms and optimise dosing regimens for clinical use.

Rosmarinus officinalis L. essential oil enhances salt stress tolerance of durum wheat seedlings through ROS detoxification and stimulation of antioxidant defense.

Salt-induced stress poses a significant barrier to agricultural productivity by impeding crop growth. Presently, environmentalists are dedicated to safeguarding food security by enhancing agricultural yields in challenging environments. Biostimulants play a crucial role in mitigating abiotic stresses in crop production, and among these, plant essential oils (EOs) stand out as organic substances with diverse biological effects on living organisms. Among the natural promoters of plant growth, Rosmarinus officinalis L. essential oil (RoEO) has gained considerable attention. Although the manifold effects of essential oils (EOs) on plant growth have been extensively demonstrated, their impact on salt stress tolerance in durum wheat seedlings remains unexplored. This investigation was undertaken to evaluate the biostimulatory capabilities of RoEO on the durum wheat cultivar "Mahmoudi." The effects of three RoEO concentrations (1, 2.5, and 5 ppm) on seed germination, growth establishment, and the induction of salt resistance under salinity conditions (150 mM NaCl) were tested. At 5 ppm, RoEO enhanced seedlings' tolerance to salinity by improving growth and reducing membrane deterioration and oxidative stress-induced damage. The expression profile analyses of seven stress-related genes (TdNHX1, TdSOS1, TdSOD, TdCAT, TdGA20-ox1, TdNRT2.1, and TdGS) using RT-qPCR showed enhancement of several important genes in durum wheat seedlings treated with 5 ppm RoEO, even under control conditions, which may be related to salt stress tolerance. The results indicate that the application of RoEO suggests a possible alternative strategy to increase salt tolerance in durum wheat seedlings towards better growth quality, thus increasing ROS scavenging and activation of antioxidant defense.

Assessment of Moroccan Cannabis Sativa Seed Oil: Chemical Analysis and Evaluation of Antioxidant, Toxicological, and Antinociceptive Effects.

Assessment of Moroccan Cannabis sativa Seed Oil: Chemical Analysis and Evaluation of Antioxidant, Toxicological, and Antinociceptive Effects. by K. Raoui et al., Cadi Ayyad University, Marrakech, Morocco. Cannabis sativa L., locally known as "El kif", belongs to the Cannabaceae family. This study aims to conduct a chemical analysis of Cannabis sativa seed oil (CSSO) and assess its acute toxicity, antioxidant properties, and analgesic effects. The chemical analysis was performed using gas chromatography and mass spectrometry (GC/MS) to identify fatty acids (FAs) contents. Antioxidant activity was evaluated in vitro using the (2,2-diphenyl-1-picrylhydrazyl) DPPH radical scavenging method and the (ferric reducing antioxidant power) FRAP method. Concurrently, acute toxicity, along with antinociceptive activity, was studied through three distinct animal models: writhing test, formalin test, and hot plate test. The results revealed that linoleic acid, oleic acid, α-linolenic acid, and palmitic acid were the main components of CSSO. The LD50 of CSSO was greater than 5 g/kg, indicating low toxicity. Additionally, CSSO exhibited a significant content of flavonoids and total polyphenols, along with notable antioxidant activity with important values. The results indicated a significant increase in thermal stimulus latency, a reduction in the number of writhes induced by acetic acid, and a decrease in licking time in both phases of the formalin test. In conclusion, this study suggests promising results for CSSO, emphasizing its potential as a therapeutic agent.

Biological Activity and Phytochemical Characteristics of Star Anise (Illicium verum) Essential Oil and Its Anti-Salmonella Activity on Sous Vide Pumpkin Model.

Illicium verum, commonly known as star anise, represents one of the notable botanical species and is recognized for its rich reservoir of diverse bioactive compounds. Beyond its culinary application as a spice, this plant has been extensively utilized in traditional medicine. Given the contemporary emphasis on incorporating natural resources into food production, particularly essential oils, to enhance sensory attributes and extend shelf life, our study seeks to elucidate the chemical composition and evaluate the antibacterial (in vitro, in situ) and insecticidal properties of Illicium verum essential oil (IVEO). Also, microbiological analyses of pumpkin sous vide treated with IVEO after inoculation of Salmonella enterica were evaluated after 1 and 7 days of study. GC/MS analysis revealed a significantly high amount of (E)-anethole (88.4%) in the investigated EO. The disc diffusion method shows that the antibacterial activity of the IVEO ranged from 5.33 (Streptococcus constellatus) to 10.33 mm (Citrobacter freundii). The lowest minimal inhibition concentration was found against E. coli and the minimum biofilm inhibition concertation was found against S. enterica. In the vapor phase, the best antimicrobial activity was found against E. coli in the pears model and against S. sonei in the beetroot model. The application of the sous vide method in combination with IVEO application decreased the number of microbial counts and eliminated the growth of S. enterica. The most isolated microbiota identified from the sous vide pumpkin were Bacillus amyloliquefaciens, B. cereus, B. licheniformis, and Ralstonia picketii. Modifications to the protein composition of biofilm-forming bacteria S. enterica were suggested by the MALDI TOF MS instigations. The IVEO showed insecticidal potential against Harmonia axyridis. Thanks to the properties of IVEO, our results suggest it can be used in the food industry as a natural supplement to extend the shelf life of foods and as a natural insecticide.

Cynara cardunculus L. var. scolymus L. Landrace "Carciofo Ortano" as a Source of Bioactive Compounds.

The preservation of agricultural biodiversity and socioeconomic development are relevant both to enhance domestic production and to support innovation. In the search for new biomolecules, we have focused on the "Carciofo Ortano" landrace, growth in the northern part of the Lazio region. Artichoke cultivation generates substantial by-products, including leaves, stems, and roots, which could serve as valuable sources of biomolecules and prebiotic dietary fiber. To valorize the leaf waste of the "Carciofo Ortano" landrace, a multidisciplinary approach was applied. Chemical analysis using HPLC-DAD identified mono-O- and di-O-caffeoylquinic acids and the sesquiterpene cynaropicrin in all artichoke leaf extracts. SPME-GC/MS analyses detected aliphatic alcohols in the fresh leaf samples. Antiproliferative and cytotoxic studies on cancer (SH-SY5Y, MCF-7, MDA) and normal (MCF-10A) human cell lines revealed that leaf extracts induced a selective dose and time-dependent biological effect. While showing slight activity against environmental bacterial strains, artichoke leaf extracts exhibited significant antifungal activity against the phytopathogenic fungus Alternaria alternata. Overall, the results highlight the potential of "Carciofo Ortano" cultivation by-products as a rich source of biomolecules with versatile applications in humans, animals, and the environment.

Changes in inoculated Salmonella enterica subsp. enterica Serovar Enteritidis and other microbiological qualities of vacuum-packed carrot slices after treatment with aqueous extract of Lobularia maritima.

After harvesting, pathogens can infect fresh vegetables in different ways. Pathogenic bacteria associated with fresh vegetables can cause widespread epidemics associated with foodborne illness. The aim of this study was to assess the microbiological quality of carrot slices after treatment with aqueous extracts of Lobularia maritima (AELm) at different concentrations AELm1 (10 mg/mL), AELm2 (5 mg/mL), AELm3 (2.5 mg/mL) and AELm4 (1.25 mg/mL), and Salmonella enterica subsp. enterica serovar Enteritidis, along with vacuum packaging and storage of carrots for 7 days at 4 °C. On days 1. and 7., total viable counts (TVC), and coliforms bacteria (CB), and Salmonella count were all analysed. Microorganisms that were obtained from carrots were identified using MALDI-TOF MS Biotyper Mass Spectrometry. The total viable, coliform bacteria and Salmonella counts were varied by the group of treatment. Higher counts were found in the control group on both days. The most isolated species of bacteria were Salmonella enterica and Pantoea agglomerans on the 1. day and Klebsiella oxytoca on the 7. day. The current study adds useful information for a better understanding of how Salmonella enterica reacts to the effect of AELm and its potential use as a sustainable washing method to eliminate bacteria from freshly cut carrots.

Chemical Investigation and Antinociceptive Activity Evaluation of Marrubium Vulgare L. Aqueous Extract.

Marrubium vulgare L. (Lamiaceae) has a long history of use in traditional herbal medicine for the treatment of respiratory tract infections, inflammatory conditions, and pain. This study aimed to investigate the chemical composition, acute toxicity, and antinociceptive effects of the aqueous extract from M. vulgare leaves (AEMV). Antioxidant activity was evaluated using DPPH and reducing power assays. The chemical composition of AEMV was determined through LC-MS/MS, and the levels of total phenolics, flavonoids, and condensed tannins were quantified. Acute oral toxicity was assessed in male Swiss mice with a single oral dose of AEMV (1, 2, 5 g/kg). The analgesic impact was examined through writhing, hot plate, and formalin tests. Our findings not only confirmed the safety of the extract in animal models but also revealed significant antioxidant activity in AEMV. High-performance liquid chromatography (HPLC) analysis identified important bioactive compounds, with marrubiin being a major component. Furthermore, AEMV demonstrated robust antinociceptive properties in all conducted tests, highlighting its potential as a valuable natural source of bioactive compounds suitable for a wide range of therapeutic applications.

Microwave-assisted hydrodiffusion and gravity extraction: An efficient method to produce pomegranate juice.

Green technologies based on microwaves have been developed by the food industry to produce organoleptically acceptable fruit juices without preliminary processing. Microwave irradiation coupled with hydrodiffusion and gravity (MHG) combines microwave heating with the earth's gravity, allowing the collection of hydrophilic substances released from the irradiated matrix. To the best of our knowledge, MHG extraction has never been experimented to produce pomegranate juice. In this work, we have evaluated it as a potential alternative to the conventional squeezing. A central composite design study (CCD) allowed the selection of the best extractive conditions (irradiation power and extraction time) to obtain a pomegranate juice with higher yield, polyphenol (e.g., catechin and delphinidin-3,5-glucoside) content, and related bioactivities (antioxidant and antidiabetic) than the one obtained by squeezing while maintaining the chemical-physical properties. Thus, this technique appears to be a functional alternative to producing high value pomegranate juice.

Molecular docking of quercetin: a promising approach for the development of new anti-inflammatory and analgesic drugs.

The aim of this study is to investigate the antinociceptive, anti-inflammatory and antipyretic effects of quercetin. Additionally, molecular docking studies were conducted to evaluate potential interactions between quercetin and various molecular targets. Animal models were used to conduct a comprehensive pharmacological investigation of quercetin. Evaluation of analgesic activity revealed a reduction in the number of abdominal cramps during the twisting test and inhibition of pain during the second phase of the formaldehyde test. Additionally, evaluation of its anti-inflammatory activity showed a reduction in ear oedema. However, it is important to note that quercetin administration has not been shown to significantly reduce yeast-induced hyperthermia. The docking study revealed the high inhibitory potential of quercetin against the COX-2 receptor.

Citrus limon Essential Oil: Chemical Composition and Selected Biological Properties Focusing on the Antimicrobial (In Vitro, In Situ), Antibiofilm, Insecticidal Activity and Preservative Effect against Salmonella enterica Inoculated in Carrot.

New goals for industry and science have led to increased awareness of food safety and healthier living in the modern era. Here, one of the challenges in food quality assurance is the presence of pathogenic microorganisms. As planktonic cells can form biofilms and go into a sessile state, microorganisms are now more resistant to broad-spectrum antibiotics. Due to their proven antibacterial properties, essential oils represent a potential option to prevent food spoilage in the search for effective natural preservatives. In this study, the chemical profile of Citrus limon essential oil (CLEO) was evaluated. GC-MS analysis revealed that limonene (60.7%), ß-pinene (12.6%), and γ-terpinene (10.3%) are common constituents of CLEO, which prompted further research on antibacterial and antibiofilm properties. Minimum inhibitory concentration (MIC) values showed that CLEO generally exhibits acceptable antibacterial properties. In addition, in situ antimicrobial research revealed that vapour-phase CLEO can arrest the growth of Candida and Y. enterocolitica species on specific food models, indicating the potential of CLEO as a preservative. The antibiofilm properties of CLEO were evaluated by MIC assays, crystal violet assays, and MALDI-TOF MS analysis against S. enterica biofilm. The results of the MIC and crystal violet assays showed that CLEO has strong antibiofilm activity. In addition, the data obtained by MALDI-TOF MS investigation showed that CLEO altered the protein profiles of the bacteria studied on glass and stainless-steel surfaces. Our study also found a positive antimicrobial effect of CLEO against S. enterica. The anti-Salmonella activity of CLEO in vacuum-packed sous vide carrot samples was slightly stronger than in controls. These results highlight the advantages of the antibacterial and antibiofilm properties of CLEO, suggesting potential applications in food preservation.

Evaluation of the Antifungal and Antiproliferative Properties of the Lichen Roccella tinctoria DC. Extracts and Main Components.

In this work, phytochemical analysis on different extracts of Roccella tinctoria DC. was reported using different techniques with respect to the past. Twenty volatile and three non-volatile compounds were identified, some of which were found in this species for the first time. The methanolic extracts and their non-volatile components were then evaluated for their antitumor effects in cancerous A549 and Mz-ChA-1 cells and for their tolerability in non-cancerous BEAS-2B and H69 cells, showing IC50 values from 94.6 µg/mL to 416.4 µg/mL, in general. The same extracts and compounds were also tested for their antifungal effects in Candida albicans, with only compound 2 being active, with an MIC50 value of 87 µg/mL. In addition, they were tested for their anti-Candida adhesion activity, anti-Candida biofilm formation, and anti-Candida mature biofilm inhibition, with efficacy percentages generally above 50% but not for all of them. Lastly, the DF3 extract and compounds 1-2 were tested in vivo according to the Galleria mellonella survival assay, showing positive mortality rates above 50% at different concentrations. All these biological assays were conducted on this species for the first time. Comparisons with other lichens and compounds were also presented and discussed.

Mixture design of α-pinene, α-terpineol, and 1,8-cineole: A multiobjective response followed by chemometric approaches to optimize the antibacterial effect against various bacteria and antioxidant activity.

α-Pinene, α-terpineol, and 1,8-cineole are compounds naturally present in essential oils, although their amounts vary from oil to oil. Although several studies have reported their antibacterial and antioxidant effects, there are few reports on the synergistic or antagonistic effects of their combinations. The objective of this study was to investigate the combined antibacterial effect of these three compounds. To our knowledge, this is the first report on the prediction of their optimal combination using the mixture design approach. The experimental antibacterial activity of the α-pinene, α-terpineol, and 1,8-cineole mixtures depended on the proportion of each compound in the mixture and the target strain, with minimum inhibitory concentrations (MIC) ranging from 0.31 to 1.85 mg/mL. Using the increased simplex-centroid mixture design, the mixture containing 0.33% of each molecule proved to be the most effective against Bacillus cereus and had the lowest MIC values. In addition, α-pinene, α-terpineol, and 1,8-cineole showed significant antioxidant activity against 2,2-picryl-1-hydrazyl radical (DPPH), with IC50 values of 24.53 ± 0.05, 65.63 ± 0.71, and 63.58 ± 0.01 µg/mL, respectively. Statistical planning and the development of utility profiles of the substance mixtures can predict the optimal composition that will exhibit the highest antibacterial activity against B. cereus as well as antioxidant properties. Furthermore, the synergistic effect of the mixtures can contribute significantly to their successful use as natural preservatives in various applications.

Long-Term Stability of Lavandula x intermedia Essential Oil Nanoemulsions: Can the Addition of the Ripening Inhibitor Impact the Biocidal Activity of the Nanoformulations?

In this work, Lavandula x intermedia essential oil (LEO) was encapsulated in lipid-based nanoemulsions (NanoLEO) using the solvent-displacement technique. In order to preserve the colloidal stability of the formulation, LEO was appropriately doped with the incorporation of different levels of a water-insoluble oil used as a ripening inhibitor. All the nanoemulsion samples were evaluated in terms of the impact of the water-insoluble oil on the nanoemulsion formation, physical-chemical properties, and antibacterial effectiveness against E. coli (Gram-negative) and B. cereus (Gram-positive). The presence of the inert oil added benefits to the formulations in terms of appearance, colloidal stability, and loss of volatile components. However, the antimicrobial activity of the nanoemulsions dramatically decreased with the ripening inhibitor addition, probably because it hampered the internalization of the antimicrobial components of LEO within the bacterial cell membranes, thus nullifying the delivery ability of the nanoemulsion formulation. On the contrary, the undoped NanoLEO formulation showed unaltered antibacterial activity in both E. coli and B. cereus up to 40 weeks from the preparation.

Anti-Salmonella Activity of Thymus serpyllum Essential Oil in Sous Vide Cook-Chill Rabbit Meat.

Food is generally prepared and vacuum-sealed in a water bath, then heated to a precise temperature and circulated in a sous vide machine. Due to its affordability and ease of use, this cooking method is becoming increasingly popular in homes and food service businesses. However, suggestions from manufacturers and chefs for long-term, low-temperature sous vide cooking raise questions about food safety in the media. In this study, heat treatment with different times and wild thyme essential oil (EO) in sous vide-processed rabbit longissimus dorsi muscle were found to inactivate Salmonella enterica. The rabbit meat samples were vacuum-packed in control groups, in the second group the rabbit meat samples were injected with S. enterica, and in the third group were meat samples infected with S. enterica with Thymus serpylum EO additive. The vacuum-packed samples were cooked sous vide for the prescribed time at 55, 60, and 65 °C. At 5, 15, 30, and 60 min, the quantities of S. enterica, total bacterial counts, and coliform bacteria were measured in groups of sous vide rabbit meat. Microbiological analyses of rabbit meat samples on days 1 and 7 were evaluated. In this study, total viable counts, coliforms bacteria, and number of Salmonella spp. were identified. After incubation, isolates from different groups of microorganisms were identified by the mass spectrometry technique. For each day measured, the test group exposed to a temperature of 55 °C for 5 min had a greater number of total microbiota. The most isolated microorganisms by MALDI-TOF MS Biotyper from the control and treated groups were Lactococcus garvieae and in the treated groups also S. enterica. Based on our analysis of sous vide rabbit meat samples, we discovered that adding 1% of thyme essential oil to the mixture reduced the amount of Salmonella cells and increased the overall and coliform bacterial counts. The microbiological quality of sous vide rabbit meat that was kept for seven days was positively impacted by the addition of thyme essential oil.

Thymus zygis, Valuable Antimicrobial (In Vitro and In Situ) and Antibiofilm Agent with Potential Antiproliferative Effects.

With the growing issues of food spoilage, microbial resistance, and high mortality caused by cancer, the aim of this study was to evaluate T. zygis essential oil (TZEO) as a potential solution for these challenges. Here, we first performed GC/MS analysis which showed that the tested TZEO belongs to the linalool chemotype since the abundance of linalool was found to be 38.0%. Antioxidant activity assays showed the superiority of TZEO in neutralizing the ABTS radical cation compared to the DPPH radical. The TZEO was able to neutralize 50% of ABTS•+ at the concentration of 53.03 ± 1.34 µg/mL. Antimicrobial assessment performed by employing disc diffusion and minimal inhibitory concentration assays revealed TZEO as a potent antimicrobial agent with the highest inhibition activity towards tested gram-negative strains. The most sensitive on the treatment with TZEO was Enterobacter aerogenes showing an MIC 50 value of 0.147 ± 0.006 mg/mL and a MIC 90 value of 0.158 ± 0.024 mg/mL. Additionally, an in situ analysis showed great effects of TZEO in inhibiting gram-negative E. coli, P. putida, and E. aerogenes growing on bananas and cucumbers. Treatment with the TZEO vapor phase in the concentration of 500 µg/mL was able to reduce the growth of these bacteria on the food models to the extent > 90%, except for E. coli growth on the cucumber, which was reduced to the extent of 83.87 ± 4.76%. Furthermore, a test on the antibiofilm activity of the tested essential oil revealed its biofilm prevention effects against Salmonella enterica which forms biofilms on plastic and stainless-steel surfaces. Performed tests on the TZEO effects towards cell viability showed no effects on the normal MRC-5 cell line. However, the results of MTT assay of TZEO effects on three cancer cell lines (MDA-MB-231, HCT-116, and K562) suggest that TZEO exerted the strongest effects on the inhibition of the viability of MDA-MB-231 cells, especially after long-term treatment in the highest concentration applied with reducing the viability of the cells to 57%. Additionally, results of NBT and Griess assays suggest that TZEO could be a convenient candidate for future testing for developing novel antitumor therapies.