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.

The Chemical Variability, Nutraceutical Value, and Food-Industry and Cosmetic Applications of Citrus Plants: A Critical Review.

Citrus fruits occupy an important position in the context of the fruit trade, considering that both fresh fruits and processed products are produced on a large scale. Citrus fruits are recognized as an essential component of the human diet, thanks to their high content of beneficial nutrients such as vitamins, minerals, terpenes, flavonoids, coumarins and dietary fibers. Among these, a wide range of positive biological activities are attributed to terpenes and flavonoids derivatives. In this review, a list of bibliographic reports (from 2015 onwards) on the phytochemical composition, beneficial effects and potential applications of citrus fruits and their by-products is systematically summarized. In detail, information regarding the nutraceutical and medicinal value closely linked to the presence of numerous bioactive metabolites and their growing use in the food industry and food packaging, also considering any technological strategies such as encapsulation to guarantee their stability over time, were evaluated. In addition, since citrus fruit, as well as its by-products, are interesting alternatives for the reformulation of natural cosmetic products, the sector of the cosmetic industry is also explored. More in-depth knowledge of the latest information in this field will contribute to future conscious use of citrus fruits.

Immunomodulatory effect of Linalool (Lin) against CCl4 -induced hepatotoxicity and oxidative damage in rats.

The current study explored the hepatoprotective and immunomodulatory effects of Linalool (Lin) against carbon tetrachloride (CCl4 )-induced toxicity in mice. Four study groups (n = 8 each) were used: (1) a negative control group and (2) a toxicity control group (single dose of CCl4 administered on day 14 as 1 mL/kg of CCL4 in 1% olive oil). Intraperitoneally (i.p.)), and two experimental groups where mice were treated with either (3) Lin (25 mg/kg b.w., orally, daily for 15 days) or (4) pretreated with Lin (25 mg/kg b.w., orally, daily for 14 days) and intoxicated with CCl4 (1 mL/kg of CCL4 in 1% olive oil. i.p.) on day 14. The levels of the anti-inflammatory cytokine interleukin 10 (IL-10), the proinflammatory cytokines TNF-α, IL-6, and TGF-1ß, and the histopathology of the liver were assessed. According to our findings, IL-10 concentrations were significantly increased in Lin-treated groups, while other cytokine levels were marked by a considerable decrease in the toxicity model group (CCl4 -treated group). Histopathological examinations of liver tissues showed that the Lin-treated groups had an almost normal structure. The current findings showed that Lin could inhibit CCl4 -induced liver injury in mice, which warrants further investigation of Lin as a potential protective and therapeutic agent against hepatotoxicity.

An In-Depth Study on the Chemical Composition and Biological Effects of Pelargonium graveolens Essential Oil.

The essential oil of Pelargonium graveolens (PGEO) is identified in the literature as a rich source of bioactive compounds with a high level of biological activity. This study aimed to examine the chemical profile of PGEO as well as its antioxidant, antibacterial, antibiofilm, and insecticidal properties. Its chemical composition was analyzed using gas chromatography-mass spectrometry (GC-MS), achieving comprehensive identification of 99.2% of volatile compounds. The predominant identified compounds were ß-citronellol (29.7%) and geraniol (14.6%). PGEO's antioxidant potential was determined by means of DPPH radical and ABTS radical cation neutralization. The results indicate a higher capacity of PGEO to neutralize the ABTS radical cation, with an IC50 value of 0.26 ± 0.02 mg/mL. Two techniques were used to assess antimicrobial activity: minimum inhibitory concentration (MIC) and disk diffusion. Antimicrobial evaluation using the disk diffusion method revealed that Salmonella enterica (14.33 ± 0.58 mm), which forms biofilms, and Priestia megaterium (14.67 ± 0.58 mm) were most susceptible to exposure to PGEO. The MIC assay demonstrated the highest performance of this EO against biofilm-forming S. enterica (MIC 50 0.57 ± 0.006; MIC 90 0.169 ± 0.08 mg/mL). In contrast to contact application, the assessment of the in situ vapor phase antibacterial activity of PGEO revealed significantly more potent effects. An analysis of antibiofilm activity using MALDI-TOF MS demonstrated PGEO's capacity to disrupt the biofilm homeostasis of S. enterica growing on plastic and stainless steel. Additionally, insecticidal evaluations indicated that treatment with PGEO at doses of 100% and 50% resulted in the complete mortality of all Harmonia axyridis individuals.

A New Approach in Meat Bio-Preservation through the Incorporation of a Heteropolysaccharide Isolated from Lobularia maritima L.

In this study, a new heteropolysaccharide extracted from Lobularia maritima (L.) Desv. (LmPS), a halophyte harvested in Tunisia, was evaluated as an antioxidant and antibacterial additive in the bio-preservation of raw minced meat. For antibacterial testing, Gram-positive bacteria such as Staphylococcus aureus ATCC and Listeria monocytogenes ATCC 19,117 and Gram-negative bacteria such as Salmonella enterica ATCC 43,972 and Escherichia coli ATCC 25,922 were used. The results indicate that this polymer had a significant antibacterial activity against foodborne pathogens. Additionally, the effects of LmPS at 0.15, 0.3 and 0.6% on refrigerated raw ground beef were investigated from a microbiological, chemical, and sensory perspective. Microbiological analysis of the meat showed that treatment with LmPS significantly (p < 0.05) improved its shelf life, while the biochemical analysis evidenced a significant (p < 0.05) decrease in lipid oxidation. LmPS at 0.6% significantly reduced by 61% and 48% metmyoglobin accumulation at the end of the storage period when compared to BHT and control samples, respectively. The chemometric approach highlighted the relationships among the different meat quality parameters. LmPS can be introduced in the food industry as a powerful natural additive and could be an alternative to synthetic antioxidant compounds.

Bio-Stimulating Effect of Natural Polysaccharides from Lobularia maritima on Durum Wheat Seedlings: Improved Plant Growth, Salt Stress Tolerance by Modulating Biochemical Responses and Ion Homeostasis.

Bioactivities of polysaccharides derived from halophyte plants have gained attention in recent years. The use of biostimulants in agriculture is an innovative method of dealing with environmental stressors affecting plant growth and development. Here, we investigated the use of natural polysaccharides derived from the halophyte plant Lobularia maritima (PSLm) as a biostimulant in durum wheat seedlings under salt stress. Treatment with polysaccharide extract (0.5, 1, and 2 mg/mL PSLm) stimulated in vitro wheat growth, including germination, shoot length, root length, and fresh weight. PSLm at 2 mg/mL provided tolerance to plants against NaCl stress with improved membrane stability and low electrolyte leakage, increased antioxidant activities (catalase (CAT), peroxidase (POD), and superoxide dismutase (SOD)), enhanced leaf chlorophyll fluorescence, proline, and total sugar contents, decreased lipid peroxidation (MDA), and reactive oxygen species (H2O2) levels, and coordinated the efflux and compartmentation of intracellular ions. The expression profile analyses of ten stress-related genes (NHX1, HKT1.4, SOS1, SOD, CAT, GA20-ox1, GA3-ox1, NRT1.1, NRT2.1, and GS) using RT-qPCR revealed the induction of several key genes in durum wheat growing in media supplemented with PSLm extract, even in unstressed conditions that could be related to the observed tolerance. This study revealed that PSLm extract contributes to salt tolerance in durum wheat seedlings, thereby enhancing their reactive oxygen species scavenging ability, and provided evidence for exploring PSLm as a plant biostimulant for sustainable and organic agriculture.

Thiamine Demonstrates Bio-Preservative and Anti-Microbial Effects in Minced Beef Meat Storage and Lipopolysaccharide (LPS)-Stimulated RAW 264.7 Macrophages.

This study assessed the anti-inflammatory effect of thiamine (TA) in lipopolysaccharide-stimulated RAW264.7 cells and also assessed the preservative properties of TA in minced beef. TA demonstrated a concentration-dependent antimicrobial effect on microbial contaminants. Inhibition zones and MIC from the effect of TA on the tested bacterial strains were respectively within the ranges 15−20 mm and 62.5−700 µg/mL. TA significantly (p < 0.05) decreased all the pro-inflammatory factors [(nitric oxide (NO), prostaglandin E2 (PGE2), TNF-α, IL-6, IL-1ß, and nuclear factor-κB (NF-κB)] monitored relative to LPS-stimulated RAW264.7 cells. TA inhibited the expression of both iNOS and COX-2. In minced beef flesh, the growth of Listeria monocytogenes was inhibited by TA. TA improved physicochemical and microbiological parameters of stored minced beef meat compared to control. Principal component analyses and heat maps elucidate the quality of the tested meats.

Insights into plant annexins function in abiotic and biotic stress tolerance.

Crop productivity depends heavily on several biotic and abiotic factors. Plant annexins are a multigene family of calcium-dependent phospholipid-binding proteins that function in response to environmental stresses and signaling during growth and development of plants. We recently isolated and characterized a Triticum durum annexin, called TdANN12, which is upregulated by different abiotic stresses. Overexpression of TdANN12 in transgenic tobacco improves stress tolerance through ROS removal. This mini-review outlines the functional characterization of plant annexin genes and suggests how these features could be exploitated to improve stress tolerance in plants. Furthermore, transgenic overexpression of plant annexin genes in crops (tobacco, tomato, rice, alfalfa, cotton, and potato) will be discussed as a promising approach to acquire abiotic and biotic stress tolerance.

Essential oil from halophyte Lobularia maritima: protective effects against CCl4-induced hepatic oxidative damage in rats and inhibition of the production of proinflammatory gene expression by lipopolysaccharide-stimulated RAW 264.7 macrophages.

The present study evaluates the chemical profiling of the essential oil of a halophyte, L. maritima (LmEO), and its protective potential against CCl4-induced oxidative stress in rats. Forty compounds have been identified in LmEO. The major components are α-pinene (3.51%), benzyl alcohol (8.65%), linalool (22.43%), pulegone (3.33%), 1-phenyl butanone (7.33%), globulol (4.32%), γ-terpinene (6.15%), terpinen-4-ol (4.31%), α-terpineol (3.9%), ledol (3.59%), epi-α-cadinol (3.05%) and α-cadinol (4.91%). In comparison with the CCl4-intoxicated group, LmEO treatment resulted in decreased liver serum marker enzymes, decreased lipid peroxidation and increased antioxidant enzyme levels, with overall further amelioration of oxidative stress. The administration of LmEO to CCl4-treated rats at a dose of 250 mg kg-1 body weight significantly reduced the toxic effects and the oxidative stress on the liver, thus validating the traditional medicinal claim of this plant. Moreover, the anti-inflammatory activity of LmEO was evaluated in lipopolysaccharide-stimulated murine RAW 264.7 cells. Our oil could modulate the inflammatory mode of the macrophages by causing reduction in iNOS and COX2 enzymes as well as in IL-1ß, IL-6, and TNF-α cytokine levels. These findings suggest that LmEO exerts anti-inflammatory effects by regulating the expression of inflammatory cytokines.

A stress-associated protein, LmSAP, from the halophyte Lobularia maritima provides tolerance to heavy metals in tobacco through increased ROS scavenging and metal detoxification processes.

Agricultural soil pollution by heavy metals is a severe global ecological problem. We recently showed that overexpression of LmSAP, a member of the stress-associated protein (SAP) gene family isolated from Lobularia maritima, in transgenic tobacco led to enhanced tolerance to abiotic stress. In this study, we characterised the response of LmSAP transgenic tobacco plants to metal stresses (cadmium (Cd), copper (Cu), manganese (Mn), and zinc (Zn)). In L. maritima, LmSAP expression increased after 12 h of treatment with these metals, suggesting its involvement in the plant response to heavy metal stress. LmSAP transgenic tobacco plants subjected to these stress conditions were healthy, experienced higher seedling survival rates, and had longer roots than non-transgenic plants (NT). However, they exhibited higher tolerance towards cadmium and manganese than towards copper and zinc. LmSAP-overexpressing tobacco seedlings accumulated more cadmium, copper, and manganese compared with NT plants, but displayed markedly decreased hydrogen peroxide (H2O2) and lipid peroxidation levels after metal treatment. Activities of the antioxidant enzymes superoxide dismutase (SOD), catalase (CAT), and peroxidase (POD) were significantly higher in transgenic plants than in NT plants after exposure to metal stress. LmSAP overexpression also enhanced the transcription of several genes encoding metallothioneins (Met1, Met2, Met3, Met4, and Met5), a copper transport protein CCH, a Cys and His-rich domain-containing protein RAR1 (Rar1), and a ubiquitin-like protein 5 (PUB1), which are involved in metal tolerance in tobacco. Our findings indicate that LmSAP overexpression in tobacco enhanced tolerance to heavy metal stress by protecting the plant cells against oxidative stress, scavenging reactive oxygen species (ROS), and decreasing the intracellular concentration of free heavy metals through its effect on metal-binding proteins in the cytosol.

Promoter of the AlSAP gene from the halophyte grass Aeluropus littoralis directs developmental-regulated, stress-inducible, and organ-specific gene expression in transgenic tobacco.

In our recent published work it has been demonstrated that AlSAP, a gene encoding an A20/AN1 zinc-finger protein (stress-associated protein) of the C4 halophyte grass Aeluropus littoralis, is inducible by various abiotic stresses and by hormonal stimuli. To further investigate the regulation of the gene, a 586-bp genomic fragment upstream of the AlSAP translated sequence has been isolated, cloned, and designated as the "Pr ( AlSAP )" promoter. Sequence analysis of "Pr ( AlSAP )" revealed the presence of cis-regulatory elements which could be required for abiotic stress, abscisic acid (ABA), and salicylic acid (SA) responsiveness and for tissue-specific and vascular expression. The Pr ( AlSAP ) promoter was fused to the ß-glucuronidase (gusA) gene and the resulting construct transferred into tobacco. Histochemical assays of stably transformed tobacco plants showed that Pr ( AlSAP ) is active in this heterologous C3 system. While full-length gusA transcripts accumulated in whole 15, 30, and 45-day-old plants, GUS histochemical staining was only observed in leaves and stems of 45-day-old, or older, transgenic seedlings. Histological sections prepared at this stage revealed activity localized in leaf veins (phloem and bundle sheath) and stems (phloem and cortex) but not in roots. Furthermore, gusA transcripts accumulated in an age-dependent manner with a basipetal pattern in leaf and stem tissues throughout the plant. In flowers, GUS expression was detected in sepals only. The accumulation of gusA transcripts was up-regulated by salt, dehydration, ABA, and SA treatment. Altogether, these results show that, when used in a heterologous dicot system, Pr ( AlSAP ) is an age-dependent, abiotic-stress-inducible, organ-specific and tissue-specific promoter.