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.

Functional Characterization of Lobularia maritima LmTrxh2 Gene Involved in Cold Tolerance in Tobacco through Alleviation of ROS Damage to the Plasma Membrane.

Cold stress is a key environmental factor affecting plant growth and development, crop productivity, and geographic distribution. Thioredoxins (Trxs) are small proteins that are ubiquitously expressed in all organisms and implicated in several cellular processes, including redox reactions. However, their role in the regulation of cold stress in the halophyte plant Lobularia maritima remains unknown. We recently showed that overexpression of LmTrxh2, which is the gene that encodes the h-type Trx protein previously isolated from L. maritima, led to an enhanced tolerance to salt and osmotic stress in transgenic tobacco. This study functionally characterized the LmTrxh2 gene via its overexpression in tobacco and explored its cold tolerance mechanisms. Results of the RT-qPCR and western blot analyses indicated differential temporal and spatial regulation of LmTrxh2 in L. maritima under cold stress at 4 °C. LmTrxh2 overexpression enhanced the cold tolerance of transgenic tobacco, as evidenced by increased germination rate, fresh weight and catalase (CAT), superoxide dismutase (SOD) and peroxidase (POD) activities; reduced malondialdehyde levels, membrane leakage, superoxide anion (O2-), and hydrogen peroxide (H2O2) levels; and higher retention of chlorophyll than in non-transgenic plants (NT). Furthermore, the transcript levels of reactive oxygen species (ROS)-related genes (NtSOD and NtCAT1), stress-responsive late embryogenis abundant protein 5 (NtLEA5), early response to dehydration 10C (NtERD10C), DRE-binding proteins 1A (NtDREB1A), and cold-responsive (COR) genes (NtCOR15A, NtCOR47, and NtKIN1) were upregulated in transgenic lines compared with those in NT plants under cold stress, indicating that LmTrxh2 conferred cold stress tolerance by enhancing the ROS scavenging ability of plants, thus enabling them to maintain membrane integrity. These results suggest that LmTrxh2 promotes cold tolerance in tobacco and provide new insight into the improvement of cold-stress resistance to cold stress in non-halophyte plants and crops.

Lobularia maritima leave extract, a nutraceutical agent with antioxidant activity, protects against CCl4-induced liver injury in mice.

Lobularia maritima (Alyssum maritimum, Brassicaceae), commonly known as sweet alyssum, is an annual ornamental halophyte widely spread along the Tunisian seashore. Lobularia maritima leaf ethanol extract was tested in an experimental model of hepatotoxicity induced by carbon tetrachloride (CCl4). L. maritima extract was found to possess in vitro antioxidant activity by scavenging the DPPH radical (IC50= 45 µg/mL), reducing/chelating iron ions and inhibiting liver lipid peroxidation induced by FeSO4. The levels of total phenolics and flavonoids were 175 ± 2.66 mg GAE/g, and 35 ± 2.88 mg QE/g respectively. Moreover, HPLC analysis revealed six compounds, namely gallic, salicylic, ellagic and ferulic acids as well as catechin and quercetin. A mice model of acute liver injury was successfully established after a single intraperitoneal injection of CCl4, as evidenced by histological analysis, Masson trichrome and Sirius red staining. Compared with the CCl4 intoxicated group, the L. maritima treatment resulted to reduce the liver serum marker enzymes, lipid peroxidation and increased the activities of antioxidant enzymes with further amelioration in the oxidative stress. The present findings discover the therapeutic potentials of L. maritima empowered with promising natural leads for the treatment of oxidative stress associated health disorders by attenuating free radicals, inhibiting lipid peroxidation, and upregulating the tissue-specific antioxidant enzymes.

Novel non-specific lipid-transfer protein (TdLTP4) isolated from durum wheat: Antimicrobial activities and anti-inflammatory properties in lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages.

Lipid transfer proteins (LTP) are members of the family of pathogenesis-related proteins (PR-14) that play a key role in plant defense mechanisms. In this study, a novel gene TdLTP4 encoding an antifungal protein from wheat (cv. Om Rabiaa) was cloned, overexpressed in Escherichia coli BL-21 (DE3) and enriched using ammonium sulfate fractionation. The TdLTP4 fusion protein was then tested against a panel of pathogens, food-borne and spoilage bacteria and fungi in order to evaluate the antimicrobial properties. TdLTP4 was applied to 0.5 µg/mL LPS-induced RAW 264.7 macrophages in vitro at different concentrations (5, 10, 20, 50 and 100 µg/mL). Levels of nitric oxide (NO), pro-inflammatory cytokines interleukin (IL)-1ß (IL-1 ß), interleukin (IL)-6 (IL-6), tumor necrosis factor (TNF-α) and anti-inflammatory cytokine IL-10 in the supernatant fraction were measured using enzyme-linked immunosorbent assay (ELISA). Expression of cyclooxygenase-2 (COX-2) and inducible nitric oxide synthase (iNOS) were detected via Western blot. The inhibition zones and minimal inhibitory concentration (MIC) values of bacterial strains were in the range of 14-26 mm and 62.5-250 µg/mL, respectively. Moreover, a remarkable activity against several fungal strains was revealed. TdLTP4 (5-100 µg/mL) decreased the production of NO (IC50 = 4.32 µg/mL), IL-6 (IC50 = 11.52 µg/mL), IL-1ß (IC50 = 7.87 µg/mL) and TNF-α (IC50 = 8.66 µg/mL) by lipopolysaccharide (LPS)-stimulated RAW 264.7 macrophages. TdLTP4 could modulate the macrophages inflammatory mode by causing reduction in iNOS and COX-2. According to these findings, TdLTP4 fusion protein could be used as natural anti-inflammatory and antimicrobial agent in food preservation and human health.

Potential anti-inflammatory and antioxidant effects of Citrus aurantium essential oil against carbon tetrachloride-mediated hepatotoxicity: A biochemical, molecular and histopathological changes in adult rats.

The present study aimed (1) to investigate the chemical composition as well as the anti-inflammatory properties and in vitro antioxidant activity of Citrus aurantium peel essential oil (pEOCa) and (2) to evaluate its potential effect in vivo. The main results showed that the major components of pEOCa are Limonene and Linalool. Additionally, DPPH scavenging ability and ß-carotene bleaching inhibition tests confirmed the antioxidant capacity of pEOCa. Our oil reduced the production of NO by LPS-stimulated RAW264,7 macrophages in a concentration-dependent. This inhibition occurred at a transcriptional level. pEOCa in CCl4 treated rats alleviated hepatotoxicity as monitored by the improvement of hepatic oxidative stress biomarkers levels plasma biochemical parameters, and DNA molecule aspect. Furthermore, the mRNA gene expression of Cu-Zn SOD, CAT, and GPx increased under CCl4 + pEOCa exposure to reach the same value to the control. Similarly, antioxidant activities of these three enzymes changed in accordance with the mRNA levels. These results were confirmed by the histological results. It seems obvious that the treatment with pEOCa prevented liver damage induced by CCl4 , thus preventing the harmful effects of free radicals.

A Brief Review on New Naturally Occurring Cembranoid Diterpene Derivatives from the Soft Corals of the Genera Sarcophyton, Sinularia, and Lobophytum Since 2016.

This work reviews the new isolated cembranoid derivatives from species of the genera Sarcophyton, Sinularia, and Lobophytum as well as their biological properties, during 2016⁻2018. The compilation permitted to conclude that much more new cembranoid diterpenes were found in the soft corals of the genus Sarcophyton than in those belonging to the genera Lobophytum or Sinularia. Beyond the chemical composition, the biological properties were also reviewed, namely anti-microbial against several Gram-positive and Gram-negative bacteria and fungi, anti-inflammatory and anti-tumoral against several types of cancer cells. In spite of the biological activities detected in almost all samples, there is a remarkable diversity in the results which may be attributed to the chemical variability that needs to be deepened in order to develop new molecules with potential application in medicine.

Phytochemical composition and antioxidant activity of Tunisian Laurus nobilis.

We study the composition of Tunisian laurel leaves essential oil (EO), the fatty acid composition of laurel seed fixed oil and the total phenolics, flavonoids and tannins of laurel leaves methanolic extract. We also evaluated its free radical scavenging activity by the DPPH test. The predominant chemical class in Tunisian Laurus nobilis leaves EO was represented by oxygenated monoterpenes accounting for 64.29% of whole EO with the major compound was 1,8-cineole (46.8%). The predominant fatty acid was oleic acid (C18: 1) with an amount of 42.0%. Total polyphenols were present in the methanolic extract of Laurusnobilis leaves at an amount of 174.1 mg GAE.g-1dry matter. Total flavonoids and total tannins accounted respectively for 149.2mg CE.g-1 dry matter and 24.9mg CEg-1 dry matter. Furthermore, concerning free radical scavenging activity, Laurus nobilis leaves methanolic extract presented a significant IC50 (3mg/mL).

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.

Enhancing Antibacterial Efficacy: Synergistic Effects of Citrus aurantium Essential Oil Mixtures against Escherichia coli for Food Preservation.

Essential oils (EOs) from various medicinal and aromatic plants are known for their diverse biological activities, including their antimicrobial effects. Citrus aurantium EO is traditionally used for therapeutic benefits due to its high content of bioactive compounds. Therefore, this study focuses on its potential use as a food preservative by investigating the combined antibacterial properties of EOs from leaves (EO1), flowers (EO2), and small branches (EO3) of Citrus aurantium against six bacterial strains by the agar disk diffusion, minimum inhibitory concentration (MIC), and minimum bactericidal concentration (MBC) methods. The chemical compositions of the EOs were analysed by gas chromatography-mass spectrometry (GC-MS) and revealed the presence of numerous compounds responsible for their antimicrobial properties. The MIC values for the EOs were 3.12 mg/mL, 4.23 mg/mL, and 1.89 mg/mL, for EO1, EO2 and EO3, respectively, while the MBC values were 12.5 mg/mL, 6.25 mg/mL, and 6.25 mg/mL, respectively. A simplex centroid design was created to analyse the effect of the individual and combined EOs against E. coli. The combined EOs showed enhanced antibacterial activity compared to the individual oils, suggesting a synergistic effect (e.g., trial 9 with an MIC of 0.21 mg/mL), allowing the use of lower EO concentrations and reducing potential negative effects on food flavour and aroma. Additionally, the practical application of investigated EOs (at concentrations twice the MIC) was investigated in raw chicken meat stored at 4 °C for 21 days. The EOs, individually and in combination, effectively extended the shelf life of the meat by inhibiting bacterial growth (total bacterial count of less than 1 × 104 CFU/g in the treated samples compared to 7 × 107 CFU/g in the control on day 21 of storage). The study underlines the potential of C. aurantium EOs as natural preservatives that represent a sustainable and effective alternative to synthetic chemicals in food preservation.

The Potential of Thymus serpyllum Essential Oil as an Antibacterial Agent against Pseudomonas aeruginosa in the Preservation of Sous Vide Red Deer Meat.

Foodborne infections caused by microbes are a serious health risk. Regarding this, customer preferences for "ready-to-eat" or minimally processed (MP) deer meat are one of the main risk factors. Given the health dangers associated with food, essential oil (EO) is a practical substitute used to decrease pathogenic germs and extend the shelf-life of MP meals. Nonetheless, further data regarding EO use in MP meals are required. In order to evaluate new, safer alternatives to chemicals for disease control and food preservation, this research was carried out in the following areas to assess the antibacterial and antibiofilm characteristics of Thymus serpyllum (TSEO) essential oil, which is extracted from dried flowering stalks. Furthermore, this study applied an essential oil of wild thyme and inoculated the sous vide deer meat with Pseudomonas aeruginosa for seven days at 4 °C in an effort to prolong its shelf-life. Against P. aeruginosa, the essential oil exhibited potent antibacterial action. The findings of the minimal biofilm inhibition concentration (MBIC) crystal violet test demonstrated the substantial antibiofilm activity of the TSEO. The TSEO modified the protein profiles of bacteria on glass and plastic surfaces, according to data from MALDI-TOF MS analysis. Moreover, it was discovered that P. aeruginosa was positively affected by the antibacterial properties of TSEO. The anti-Pseudomonas activity of the TSEO was marginally higher in vacuum-packed sous vide red deer meat samples than in control samples. The most frequently isolated species from sous vide deer meat, if we do not consider the applied bacteria Pseudomonas aeruginosa, were P. fragi, P. lundensis, and P. taetrolens. These results highlight the antibacterial and antibiofilm qualities of TSEO, demonstrating its potential for food preservation and extending the shelf-life of deer meat.

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.

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.

Insight into Pelargonium odoratissimum Essential Oil Preservative Properties Effect on Ground Beef.

Pelargonium plants are very popular and well-known for their essential oils (EOs), which are used for medicinal purposes and in food. This study focused on the EO of Pelargonium odoratissimum. First, its composition and antioxidant and antimicrobial activity were evaluated, and finally, its efficacy as a natural preservative in ground beef was tested. The main EO constituents were citronellol (40.0%), nerol (15.3%), and citronellyl formate (12.6%). The antibacterial activity of POEO showed that Enterococcus faecalis ATCC 29212 was the most susceptible strain compared to the other eight strains tested. The antioxidant activity, as measured by the DPPH assay, showed a dose-dependent effect with an IC50 comparable to the standard used, gallic acid. Aerobic plate count, psychotropic bacteria, and Enterobacteriaceae, including Salmonella, were reduced by the addition of Pelargonium odoratissimum essential oils. The oxidative stability was significantly improved compared to the untreated sample. Additionally, the results for metmyoglobin demonstrated a notable preservative effect on sensory properties, including appearance, odor, color, and overall acceptability. The ability to discriminate between all samples and correlate protein and lipid oxidation processes, microbiological characteristics, and sensory measurements was made possible by principal component analysis and heat maps. This research shows the potential benefits of using POEO in the preservation of ground beef by effectively extending shelf life and improving product safety.

The interaction between formylphenoxyacetic acid derivatives (chalcone and flavones) and ionic surfactants: Insights into binding constants, solubilisation and physiochemical properties.

In this study, UV-vis spectroscopy was employed to investigate the interaction between formylphenoxyacetic acid (FPAA) and its derivatives (chalcone and flavones) with ionic surfactants (SDS, CTAB, and DTAB) in different physiological environments. Changes in the physiochemical properties of FPAA chalcone and flavones including binding constants, partitioning constants, and Gibbs free energy were observed which were influenced by the presence of ionic surfactants computed using mathematical models. The solubilization of the targeted compounds in the ionic surfactants was determined through the binding constant (Kb). The results of the present study indicated that electrostatic interactions played a significant role in the solubilization of the targeted compounds in SDS, CTAB, and DTAB. At pH 4.1, FPAA chalcone exhibited stronger binding affinity with SDS compared to CTAB and DTAB. However, at pH 7.4, chalcone showed stronger binding with DTAB compared to SDS, while negligible interaction with CTAB was observed at pH 7.4. The flavones demonstrated stronger binding with DTAB at pH 7.4 compared to SDS and CTAB and it exhibited strong bonding with CTAB at pH 4.1. The negative values of the Gibbs free energy for binding (ΔGb˚) and partitioning (ΔGp˚) constants displayed the spontaneity of the process. However, FPAA chalcone with SDS and FPAA flavones with DTAB furnished positive ΔGb˚, indicating a non-spontaneous process.

Fatty Acid Profile and Genetic Variants of Proteins Involved in Fatty Acid Metabolism Could Be Considered as Disease Predictor.

Circulating fatty acids (FA) have an endogenous or exogenous origin and are metabolized under the effect of many enzymes. They play crucial roles in many mechanisms: cell signaling, modulation of gene expression, etc., which leads to the hypothesis that their perturbation could be the cause of disease development. FA in erythrocytes and plasma rather than dietary FA could be used as a biomarker for many diseases. Cardiovascular disease was associated with elevated trans FA and decreased DHA and EPA. Increased arachidonic acid and decreased Docosahexaenoic Acids (DHA) were associated with Alzheimer's disease. Low Arachidonic acid and DHA are associated with neonatal morbidities and mortality. Decreased saturated fatty acids (SFA), increased monounsaturated FA (MUFA) and polyunsaturated FA (PUFA) (C18:2 n-6 and C20:3 n-6) are associated with cancer. Additionally, genetic polymorphisms in genes coding for enzymes implicated in FA metabolism are associated with disease development. FA desaturase (FADS1 and FADS2) polymorphisms are associated with Alzheimer's disease, Acute Coronary Syndrome, Autism spectrum disorder and obesity. Polymorphisms in FA elongase (ELOVL2) are associated with Alzheimer's disease, Autism spectrum disorder and obesity. FA-binding protein polymorphism is associated with dyslipidemia, type 2 diabetes, metabolic syndrome, obesity, hypertension, non-alcoholic fatty liver disease, peripheral atherosclerosis combined with type 2 diabetes and polycystic ovary syndrome. Acetyl-coenzyme A carboxylase polymorphisms are associated with diabetes, obesity and diabetic nephropathy. FA profile and genetic variants of proteins implicated in FA metabolism could be considered as disease biomarkers and may help with the prevention and management of diseases.

Salvia officinalis L. and Salvia sclarea Essential Oils: Chemical Composition, Biological Activities and Preservative Effects against Listeria monocytogenes Inoculated into Minced Beef Meat.

In this study, Salvia officinalis L. and Salvia sclarea essential oils (EOs) were investigated using gas chromatography-mass spectrometry (GC-MS) to describe their chemical composition. The obtained results show, for both EOs, a profile rich in terpene metabolites, with monoterpenes predominating sesquiterpenes but with significant qualitative and quantitative differences. The main compound found in the Salvia officinalis EO (SOEO) was camphor (19.0%), while in Salvia sclarea EO (SCEO), it was linalyl acetate (59.3%). Subsequently, the in vitro antimicrobial activity of the EOs against eight pathogenic strains was evaluated. The disc diffusion method showed a significant lysis zone against Gram-positive bacteria. The minimum inhibitory concentrations (MICs) ranged from 3.7 mg/mL to 11.2 mg/mL, indicating that each EO has specific antimicrobial activity. Both EOs also showed significant antiradical activity against DPPH radicals and total antioxidant activity. In addition, the preservative effect of SOEO (9.2%) and SCEO (9.2%), alone or in combination, was tested in ground beef, and the inhibitory effect against Listeria monocytogenes inoculated into the raw ground beef during cold storage was evaluated. Although the effect of each individual EO improved the biochemical, microbiological, and sensory parameters of the samples, their combination was more effective and showed complete inhibition of L. monocytogenes after 7 days of storage at 4 °C. The results show that both EOs could be used as safe and natural preservatives in various food and/or pharmaceutical products.

Biopreservative Effect of the Tunisian Halophyte Lobularia maritima Flavonoid Fraction, Used Alone and in Combination with Linalool in Stored Minced Beef Meat.

In the present study, Lobularia maritima (Lm) flavonoid extract (LmFV) was characterized by HPLC analyses and five compounds were detected. Further, to describe the chemical content of the matrix, GC-MS analyses after silylation were performed; the obtained results showed the presence of a large number of components belonging to several chemical classes, mostly sugar alcohols, sugars, fatty acids, and terpenes. Firstly, the antibacterial activities of this fraction and linalool (Lin) were evaluated against eight foodborne pathogenic strains with MIC values between 2.3 and 5.8 mg/mL and 0.23 and 0.7 mg/mL, respectively. Then, the antioxidant activity of both was evaluated by the DPPH antiradical test and the phosphomolybdenum test. Furthermore, the biopreservative effect of LmFV alone and in combination with Lin on minced beef stored at 4 °C for 14 days was evaluated using microbiological and physiochemical tests. LmFV at 4.6% alone significantly reduced microbial spoilage in ground meat (p < 0.05). The combination of LmFV (4.6%) and Lin (0.46%) was more effective than LmFV alone in inhibiting bacterial contamination, reducing TBARS values and the risk of bacterial contamination, and reducing the accumulation of Met myoglobin (MetMb). This combination, therefore, extends the shelf life of the product by about 10 days. Based on these microbiological results and physicochemical parameters, it can be stated that the addition of Lin potentiates the flavonoid fraction of L. maritima more strongly against the deterioration of meat quality by significantly improving its biopreservative effect as a natural conservative.

High Biofilm-Forming Ability and Clonal Dissemination among Colistin-Resistant Escherichia coli Isolates Recovered from Cows with Mastitis, Diarrheic Calves, and Chickens with Colibacillosis in Tunisia.

BACKGROUND: Escherichia coli (E. coli) is one of the main etiological agents responsible for bovine mastitis (BM), neonatal calf diarrhea (NCD), and avian colibacillosis (AC). This study aimed to assess resistance and virulence genes content, biofilm-forming ability, phylogenetic groups, and genetic relatedness in E. coli isolates recovered from clinical cases of BM, NCD, and AC. MATERIALS/METHODS: A total of 120 samples including samples of milk (n = 70) and feces (n = 50) from cows with BM and calves with NCD, respectively, were collected from different farms in Northern Tunisia. Bacterial isolation and identification were performed. Then, E. coli isolates were examined by disk diffusion and broth microdilution method for their antimicrobial susceptibility and biofilm-forming ability. PCR was used to detect antimicrobial resistance genes (ARGs), virulence genes (VGs), phylogenetic groups, and Enterobacterial repetitive intergenic consensus PCR (ERIC-PCR) for their clonal relationship. RESULTS: Among the 120 samples, 67 E. coli isolates (25 from BM, 22 from AC, and 20 from NCD) were collected. Overall, 83.6% of isolates were multidrug resistant. Thirty-six (53.73%) isolates were phenotypically colistin-resistant (CREC), 28.3% (19/67) were ESBL producers (ESBL-EC), and forty-nine (73.1%) formed biofilm. The blaTEM gene was found in 73.7% (14/19) of isolates from the three diseases, whilst the blaCTXM-g-1 gene was detected in 47.3% (9/19) of isolates, all from AC. The most common VG was the fimA gene (26/36, 72.2%), followed by aer (12/36, 33.3%), cnf1 (6/36, 16.6%), papC (4/36, 11.1%), and stx1 and stx2 genes (2/36; 5.5% for each). Phylogenetic analysis showed that isolates belonged to three groups: A (20/36; 55.5%), B2 (7/36; 19.4%), and D (6/36; 16.6%). Molecular typing by ERIC-PCR showed high genetic diversity of CREC and ESBL E. coli isolates from the three animal diseases and gave evidence of their clonal dissemination within farms in Tunisia. CONCLUSION: The present study sheds new light on the biofilm-forming ability and clonality within CREC and ESBL-EC isolated from three different animal diseases in Tunisian farm animals.

Recent Advances in the Incorporation of Polysaccharides with Antioxidant and Antibacterial Functions to Preserve the Quality and Shelf Life of Meat Products.

Meat and meat products are susceptible to various types of natural processes such as oxidative degradation due to their high content of protein and essential amino acids. However, finding solutions to maintain the nutritional and sensory quality of meat and meat products is unavoidable. Hence, there is a pressing need to investigate alternatives to synthetic preservatives, focusing on active biomolecules of natural provenance. Polysaccharides are natural polymers of various sources that exhibit antibacterial and antioxidant properties via a variety of mechanisms, owing to their diversity and structural variation. For this reason, these biomolecules are widely studied in order to improve texture, inhibit the growth of pathogens, and improve the oxidative stability and sensory characteristics of meat products. However, the literature has not addressed their biological activity in meat and meat products. This review summarizes the various sources of polysaccharides, their antioxidant and antibacterial activities (mainly against pathogenic food strains), and their use as natural preservatives to replace synthetic additives in meat and meat products. Special attention is given to the use of polysaccharides to improve the nutritional value of meat, resulting in more nutrient-rich meat products with higher polysaccharide content and less salt, nitrites/nitrates, and cholesterol.

Dried Herbs as an Easy-to-Use and Cost-Effective Alternative to Essential Oils to Extend the Shelf Life of Sheep Lump Cheese.

The objective of this research was to assess the effectiveness of three specific dried herbs (rosemary, thyme, and oregano) in combating microbial spoilage in sheep lump cheese. This was achieved by comparing them with a control group and cheeses treated with corresponding 1% essential oils (Rosmarinus officinalis, Origanum vulgare, Thymus vulgaris). All cheese samples were vacuum-sealed and stored at 4 °C for 15 days. Analysis of total viable counts of viable bacteria (TVC), coliform bacteria (CB), lactic acid bacteria (LAB), and microscopic filamentous fungi (MFF) was conducted on days 0, 5, 10, and 15. The results revealed that, at the end of the storage period, dried oregano-treated samples exhibited the lowest TVC count (5.80 log CFU/g), while dried rosemary-treated samples showed the lowest CB count (3.27 log CFU/g). Moreover, the lowest MFF count (2.40 log CFU/g) was observed in oregano essential oil-treated samples. Additionally, dried oregano-treated samples displayed the highest LAB count (4.49 log CFU/g) at the experiment's conclusion. Furthermore, microorganism identification from sheep cheese was performed using MALDI-TOF MS Biotyper technology, revealing that the most frequently isolated bacteria were Citrobacter braakii and Hafnia alvei (Enterobacteriaceae family), along with Lacticaseibacillus paracasei (Lactobacillaceae family). In summary, all the natural substances examined exhibited inhibitory effects against the studied microorganisms, with oregano essential oil and dried oregano demonstrating the strongest inhibitory effects. This supports their potential use as cost-effective natural preservatives to extend the shelf life of sheep lump cheese.