Allethrin Promotes Apoptosis and Autophagy Associated with the Oxidative Stress-Related PI3K/AKT/mTOR Signaling Pathway in Developing Rat Ovaries.

The increased concern regarding the reduction in female fertility and the impressive numbers of women undergoing fertility treatment support the existence of environmental factors beyond inappropriate programming of developing ovaries. Among these factors are pyrethroids, which are currently some of the most commonly used pesticides worldwide. The present study was performed to investigate the developmental effects of the pyrethroid-based insecticide allethrin on ovarian function in rat offspring in adulthood. We mainly focused on the roles of oxidative stress, apoptosis, autophagy and the related pathways in ovarian injury. Thirty-day-old Wistar albino female rats were intragastrically administered 0 (control), 34.2 or 68.5 mg/kg body weight allethrin after breeding from Day 6 of pregnancy until delivery. We found that allethrin-induced ovarian histopathological damage was accompanied by elevations in oxidative stress and apoptosis. Interestingly, the number of autophagosomes in allethrin-treated ovaries was higher, and this increase was correlated with the upregulated expression of genes and proteins related to the autophagic marker LC-3. Furthermore, allethrin downregulated the expression of PI3K, AKT and mTOR in allethrin-treated ovaries compared with control ovaries. Taken together, the findings of this study suggest that exposure to the pyrethroid-based insecticide allethrin adversely affects both the follicle structure and function in rat offspring during adulthood. Specifically, allethrin can induce excessive oxidative stress and defective autophagy-related apoptosis, probably through inactivation of the PI3K/AKT/mTOR signaling pathway, and these effects may contribute to ovarian dysfunction and impaired fertility in female offspring.

An agar-based bioassay for accurate screening of the total antioxidant capacity of lactic acid bacteria cell-free supernatants.

This study aims to develop a new, simple, and efficient method for estimating the total antioxidant capacity of lactic acid bacteria-cell free supernatant. The bioassay is based on diffusion and reduction of permanganate in agar medium under acidic conditions where the Mn7+ ions are quantitatively oxidized to Mn2+ and shift from an intense purple color to colorless. Hence, the reaction enables fast detection of the bleaching diameter during diffusion of the sample in permanganate agar. This bleaching diameter is correlated to the reducing power of the substance tested. The method was tested and validated to quantify the total antioxidant capacity of culture supernatants of probiotic strains (Lacticaseibacillus rhamnosus LGG and Lactiplantibacillus plantarum 299v) and 25 lactic acid bacteria isolated from a human intestinal origin and compared to the PRAC and DPPH methods. The results were treated statistically by analysis of variance. This method proved to be linear (R2 in the linear experiment of ascorbic acid was 0,99), precise with repeatability intraday RSD of 2.07 to 5.5% and intermediate precision RSD of 2.95 to 5.53%, and accurate (100.29 to 108.58%) at 30 min, 1 h, and 4 h in the selected range of 1.5-5.5 mM of ascorbic acid. The developed permanganate agar reduction bioassay is a fast, reliable, and cost-effective technique for the prescreening and detecting the total antioxidant capacity of supernatants of lactic acid bacteria and possibly other sources of natural antioxidants.

Enterococcus spp.: Is It a Bad Choice for a Good Use-A Conundrum to Solve?

Since antiquity, the ubiquitous lactic acid bacteria (LAB) Enterococci, which are just as predominant in both human and animal intestinal commensal flora, have been used (and still are) as probiotics in food and feed production. Their qualities encounter several hurdles, particularly in terms of the array of virulence determinants, reflecting a notorious reputation that nearly prevents their use as probiotics. Additionally, representatives of the Enterococcus spp. genus showed intrinsic resistance to several antimicrobial agents, and flexibility to acquire resistance determinants encoded on a broad array of conjugative plasmids, transposons, and bacteriophages. The presence of such pathogenic aspects among some species represents a critical barrier compromising their use as probiotics in food. Thus, the genus neither has Generally Recognized as Safe (GRAS) status nor has it been included in the Qualified Presumption of Safety (QPS) list implying drastic legislation towards these microorganisms. To date, the knowledge of the virulence factors and the genetic structure of foodborne enterococcal strains is rather limited. Although enterococcal infections originating from food have never been reported, the consumption of food carrying virulence enterococci seems to be a risky path of transfer, and hence, it renders them poor choices as probiotics. Auspiciously, enterococcal virulence factors seem to be strain specific suggesting that clinical isolates carry much more determinants that food isolates. The latter remain widely susceptible to clinically relevant antibiotics and subsequently, have a lower potential for pathogenicity. In terms of the ideal enterococcal candidate, selected strains deemed for use in foods should not possess any virulence genes and should be susceptible to clinically relevant antibiotics. Overall, implementation of an appropriate risk/benefit analysis, in addition to the case-by-case assessment, the establishment of a strain's innocuity, and consideration for relevant guidelines, legislation, and regulatory aspects surrounding functional food development seem to be the crucial elements for industries, health-staff and consumers to accept enterococci, like other LAB, as important candidates for useful and beneficial applications in food industry and food biotechnology. The present review aims at shedding light on the world of hurdles and limitations that hampers the Enterococcus spp. genus and its representatives from being used or proposed for use as probiotics. The future of enterococci use as probiotics and legislation in this field are also discussed.

Draft Genome Sequences of Five Potentially Probiotic Enterococcus faecium Strains Isolated from an Artisanal Tunisian Meat (Dried Ossban).

Here, we report the first draft genome sequences of five bacteriocinogenic and potentially probiotic Enterococcus faecium strains (MZF1 to MZF5), which were isolated from homemade Tunisian meat (dried ossban). The estimated median genome sizes were about 2,582,641 ± 109,039 bp, with a median G+C content of 40% ± 0.4%.

In vitro Assessment of the Probiotic Properties and Bacteriocinogenic Potential of Pediococcus pentosaceus MZF16 Isolated From Artisanal Tunisian Meat "Dried Ossban".

Pediococcus pentosaceus MZF16 has been isolated from artisanal Tunisian meat so called "Dried Ossban," an original ecological niche, and identified by MALDI-TOF mass spectrometry and 16S rDNA sequencing. This bacterium showed a high tolerance to gastric stress conditions, and toward bile salts. P. pentosaceus MZF16 also demonstrated a hydrophobic surface profile (high adhesion to xylene), autoaggregation, and adhesive abilities to the human intestinal Caco-2/TC7 cell line. These properties may help the bacterium colonizing the gut. Furthermore, MZF16 was found to be resistant to gentamycin and chloramphenicol but did not harbor any transferable resistance determinants and/or virulence genes. The data also demonstrated absence of cytotoxicity of this strain. Conversely, P. pentosaceus MZF16 can slightly stimulate the immune system and enhance the intestinal epithelial barrier function. Moreover, this bacterium has been shown to be highly active against Listeria spp. due to bacteriocin production. Characterization of the bacteriocin by PCR amplification, sequencing and bioinformatic analyses revealed that MZF16 produces a bacteriocin 100% identical to coagulin, a pediocin-like inhibitory substance produced by Bacillus coagulans. To our knowledge, this is the first report that highlights the production of a pediocin 100% identical to coagulin in a Pediococcus strain. As coagulin, pediocin MZF16 has the consensus sequence YYGNGVXCXXXXCXVXXXXA (X denotes any amino acid), which confirms its belonging to class IIa bacteriocins, and its suitability to preserve foods from Listeria monocytogenes development. According to these results, P. pentosaceus MZF16 can be proposed as a probiotic and bioprotective agent for fermented foods, including Tunisian dry meat and sausages. Further investigations will aim to study the behavior of this strain in meat products as a component of functional food.

Evaluation of Probiotic Properties and Safety of Enterococcus faecium Isolated From Artisanal Tunisian Meat "Dried Ossban".

Enterococcus faecium strains were isolated from an original biotope, artisanal dried Tunisian meat "Dried Ossban," and evaluated for safety and capacity as probiotics. Gram-positive, catalase negative, and bacteriocin-producing bacteria were screened using selective microbiological media. All isolates were identified by phenotypic and molecular tools. Five E. faecium strains (MZF1, MZF2, MZF3, MZF4, and MZF5) were selected and further assessed for their probiotic properties. They were found to be resistant to the physiological concentrations of bile salts, and the harsh conditions of the gastrointestinal tract, and showed autoaggregation and adhesion ability. All these isolates possess at least one enterocin and could efficiently inhibit the growth of Listeria innocua HPB13. The analysis of their safety profile revealed for almost all the strains the absence of cytotoxicity and virulence determinants, and susceptibility to clinically important antibiotics such as vancomycin. These data suggest that these bacteria, isolated from "Dried Ossban," do not present a risk to human health, and may be considered as interesting candidates for future use as probiotics and bioprotective cultures for application in the food and/or feed industries.

The Genus Enterococcus: Between Probiotic Potential and Safety Concerns-An Update.

A considerable number of strains belonging to different species of Enterococcus are highly competitive due to their resistance to wide range of pH and temperature. Their competitiveness is also owed to their ability to produce bacteriocins recognized for their wide-range effectiveness on pathogenic and spoilage bacteria. Enterococcal bacteriocins have attracted great research interest as natural antimicrobial agents in the food industry, and as a potential drug candidate for replacing antibiotics in order to treat multiple drugs resistance pathogens. However, the prevalence of virulence factors and antibiotic-resistance genes and the ability to cause disease could compromise their application in food, human and animal health. From the current regulatory point of view, the genus Enterococcus is neither recommended for the QPS list nor have GRAS status. Although recent advances in molecular biology and the recommended methods for the safety evaluation of Enterococcus strains allowed the distinction between commensal and clinical clades, development of highly adapted methods and legislations are still required. In the present review, we evaluate some aspects of Enterococcus spp. related to their probiotic properties and safety concerns as well as the current and potential application in food systems and treatment of infections. The regulatory status of commensal Enterococcus candidates for food, feed, probiotic use, and recommended methods to assess and ensure their safety are also discussed.

ANTISTAPHYBASE: database of antimicrobial peptides (AMPs) and essential oils (EOs) against methicillin-resistant Staphylococcus aureus (MRSA) and Staphylococcus aureus.

Staphylococcus aureus and methicillin-resistant S. aureus are major pathogens. The antimicrobial peptides and essential oils (EOs) display narrow- or broad-spectrum activity against bacteria including these strains. A centralized resource, such as a database, designed specifically for anti-S. aureus/anti-methicillin-resistant S. aureus antimicrobial peptides and EOs is therefore needed to facilitate the comprehensive investigation of their structure/activity associations and combinations. The database ANTISTAPHYBASE is created to facilitate access to important information on antimicrobial peptides and essential peptides against methicillin-resistant S. aureus and S. aureus. At the moment, the database contains 596 sequences of antimicrobial peptides produced by diverse organisms and 287 essential oil records. It permits a quick and easy search of peptides based on their activity as well as their general, physicochemical properties and literature data. These data are very useful to perform further bioinformatic or chemometric analysis and would certainly be useful for the development of new drugs for medical use. The ANTISTAPHYBASE database is freely available at: https://www.antistaphybase.com/ .

Inhibition of methicillin-resistant Staphylococcus aureus (MRSA) by antimicrobial peptides (AMPs) and plant essential oils.

CONTEXT: Drug-resistant bacterial infections cause considerable patient mortality and morbidity. The annual frequency of deaths from methicillin-resistant Staphylococcus aureus (MRSA) has surpassed those caused by human immunodeficiency virus/acquired immune deficiency syndrome. The antimicrobial peptides (AMPs), plant essential oils (EOs) and their combinations have proven to be quite effective in killing a wide selection of bacterial pathogens including MRSA. OBJECTIVES: This review summarizes the studies in the use of AMPs, plant EOs and their combinations for coping with MRSA bacteria, and to formulate new prospects for future studies on this topic. METHODS: The sources of scientific literature such as PubMed, library search, Google Scholar, Science Direct and electronic databases such as 'The Antimicrobial Peptide Database', 'Collection of Anti-Microbial Peptides' and 'YADAMP'. Physicochemical data of anti-MRSA peptides were determined by Scientific DataBase Maker software. RESULTS: Of the 118 peptides, 88 exhibited an activity against MRSA with the highest activity of minimum inhibitory concentration values. Various plant EOs have been effective against MRSA. Remarkably, lemongrass EOs completely inhibited all MRSA growth on the plate. Lemon myrtle, Mountain savory, Cinnamon bark and Melissa EOs showed a significant inhibition. CONCLUSION: Several of these AMPs, EOs and their combinations were effective against MRSA. Their activities have implications for the development of new drugs for medical use.

Chemical composition and antibacterial activities of seven Eucalyptus species essential oils leaves.

BACKGROUND: In this paper, we have studied the essential oils chemical composition of the leaves of seven Eucalyptus species developed in Tunisia. Eucalyptus leaves were picked from trees growing in different arboretums in Tunisia. Choucha and Mrifeg arboretums located in Sedjnene, region of Bizerte (Choucha: E. maideni, E. astrengens et E. cinerea; Mrifeg : E. leucoxylon), Korbous arboretums located in the region of Nabeul, North East Tunisia with sub-humid bioclimate, (E. lehmani), Souiniet-Ain Drahem arboretum located in region of Jendouba (E. sideroxylon, E. bicostata). Essential oils were individually tested against a large panel of microorganisms including Staphylococcus aureus (ATCC 6539), Escherichia coli (ATCC 25922), Enterococcus faecalis (ATCC29212), Listeria ivanovii (RBL 30), Bacillus cereus (ATCC11778). RESULTS: The yield of essential oils ranged from 1.2% to 3% (w/w) for the different Eucalyptus species. All essential oils contain α-pinene, 1,8-cineol and pinocarveol-trans for all Eucalyptus species studied. The 1,8-cineol was the major compound in all species (49.07 to 83.59%). Diameter of inhibition zone of essential oils of Eucalyptus species varied from 10 to 29 mm. The largest zone of inhibition was obtained for Bacillus cereus (E. astrengens) and the lowest for Staphylococcus aureus (E. cinerea). The essential oils from E. maideni, E. astrengens, E. cinerea (arboretum of Bizerte), E. bicostata (arboretum of Aindraham) showed the highest antibacterial activity against Listeria ivanovii and Bacillus cereus. CONCLUSION: The major constituents of Eucalyptus leaves essential oils are 1,8-cineol (49.07 to 83.59%) and α-pinene (1.27 to 26.35%). The essential oils from E. maideni, E. astrengens, E. cinerea, E. bicostata showed the highest antibacterial activity against Listeria ivanovii and Bacillus cereus, they may have potential applications in food and pharmaceutical products.

Chemical composition and antibacterial activities of seven Eucalyptus species essential oils leaves

BACKGROUND: In this paper, we have studied the essential oils chemical composition of the leaves of seven Eucalyptus species developed in Tunisia. Eucalyptus leaves were picked from trees growing in different arboretums in Tunisia. Choucha and Mrifeg arboretums located in Sedjnene, region of Bizerte (Choucha: E. maideni, E. astrengens et E. cinerea; Mrifeg : E. leucoxylon), Korbous arboretums located in the region of Nabeul, North East Tunisia with sub-humid bioclimate, (E. lehmani), Souiniet-Ain Drahem arboretum located in region of Jendouba (E. sideroxylon, E. bicostata). Essential oils were individually tested against a large panel of microorganisms includingStaphylococcus aureus (ATCC 6539), Escherichia coli (ATCC 25922), Enterococcus faecalis (ATCC29212), Listeria ivanovii (RBL 30), Bacillus cereus (ATCC11778). RESULTS: The yield of essential oils ranged from 1.2% to 3% (w/w) for the different Eucalyptus species. All essential oils contain α-pinene, 1,8-cineol and pinocarveol-trans for all Eucalyptus species studied. The 1,8-cineol was the major compound in all species (49.07 to 83.59%). Diameter of inhibition zone of essential oils of Eucalyptus species varied from 10 to 29 mm. The largest zone of inhibition was obtained for Bacillus cereus (E. astrengens) and the lowest for Staphylococcus aureus (E. cinerea). The essential oils from E. maideni, E. astrengens, E. cinerea (arboretum of Bizerte), E. bicostata(arboretum of Aindraham) showed the highest antibacterial activity against Listeria ivanovii and Bacillus cereus. CONCLUSION: The major constituents of Eucalyptus leaves essential oils are 1,8-cineol (49.07 to 83.59%) and α-pinene (1.27 to 26.35%). The essential oils from E. maideni, E. astrengens, E. cinerea, E. bicostatashowed the highest antibacterial activity against Listeria ivanovii and Bacillus cereus, they may have potential applications in food and pharmaceutical products.

Flaxseed hull: Chemical composition and antioxidant activity during development.

Changes in the chemical composition and antioxidant activity of flaxseed hull during maturation were investigated. P129 hull variety was studied at four maturation stages (St1, St2, St3, and St4). Significant variation in proximate composition and flaxseed hull oil characteristics were observed. A significant increase in the carbohydrates content of the hull was observed during development. The main methyl esters were linolenic acid (48.95 - 51.52 %), oleic acid (20.27-23.41%) and linoleic acid (15.62-17.70%). The highest polyunsaturated fatty acids (PUFA) were found to be 67.14 % at the first stage of maturity (St1). Flaxseed hull oil was of good quality, containing an abundance of omega-3 essential fatty acids. The iodine value increased, while the saponification value of oil decreased during seed development. The decrease in ascorbic acid content was steady. The maximum level of total phenolic acid content (128.3 mg/100 g oil) was reached at 7 DAF. The antioxidant activity of oilseed was assessed by means of 2, 2-diphenyl-1-picrylhydrazyl (DPPH) radical-scavenging assay. Radical scavenging activity for green hull was 52.74% and mature hull was 69.32%.

Lipids, proteins, phenolic composition, antioxidant and antibacterial activities of seeds of peanuts (Arachis hypogaea l) cultivated in Tunisia.

Fatty acid composition of peanut seed oil in four varieties cultivated in Tunisia showed that linoleic (C18:2), oleic (C18:1) and palmitic (C16) acids account for more than 84% for Chounfakhi and Massriya and for more than 85% of the total fatty acids of Trabilsia and Sinya seed oil respectively. Seed oil contents were significantly different (P ≤ 0.05) and did not exceed 48%. The study of total phenolics revealed that Chounfakhi contained more total phenolics (2.1 mg GAE/g DW), followed by the Massriya and Sinya cultivars (1.35 mg GAE/g DW for each); Trabilsia presented the lowest total phenolic content with 1 mg GAE/g DW. Considerable antiradical ability was found, especially in the Trabilsia peanut seed cultivar (IC50 = 1550 µg/ml), the Massriya and Sinya cultivars had, respectively, 720 and 820 mg/ml IC50. In the Massriya variety the sterol fraction showed antibacterial activity against Listeria ivanovii, Listeria inocua, Pseudomonas aeruginosa, Staphylococus aureus, Enterococcus hirae and Bacillus cereus.

Lipids, proteins, phenolic composition, antioxidant and antibacterial activities of seeds of peanuts (Arachis hypogaea l) cultivated in Tunisia

Fatty acid composition of peanut seed oil in four varieties cultivated in Tunisia showed that linoleic (C18:2), oleic (C18:1) and palmitic (C16) acids account for more than 84% for Chounfakhi and Massriya and for more than 85% of the total fatty acids of Trabilsia and Sinya seed oil respectively. Seed oil contents were significantly different (P ≤ 0.05) and did not exceed 48%. The study of total phenolics revealed that Chounfakhi contained more total phenolics (2.1 mg GAE/g DW), followed by the Massriya and Sinya cultivars (1.35 mg GAE/g DW for each); Trabilsia presented the lowest total phenolic content with 1 mg GAE/g DW. Considerable antiradical ability was found, especially in the Trabilsia peanut seed cultivar (IC50 = 1550 μg/ml), the Massriya and Sinya cultivars had, respectively, 720 and 820 mg/ml IC50. In the Massriya variety the sterol fraction showed antibacterial activity against Listeria ivanovii, Listeria inocua, Pseudomonas aeruginosa, Staphylococus aureus, Enterococcus hirae and Bacillus cereus.

Phytosterols accumulation in the seeds of Linum usitatissimum L.

A comparative study was performed to determine the free sterols content and composition during the development of three varieties of linseed (H52, O116 and P129). Seed samples were collected at regular intervals from 7 to 60 days after flowering (DAF). Ten compounds were identified: cholesterol, campesterol, brassicasterol, stigmasterol, beta-sitosterol, Delta5-avenasterol, cycloartenol; 24-methylene cycloartanol, obtusifoliol, citrostadienol. The maximum level of 4-desmethylsterols (1,515 mg/100g oil) was reached at 7 DAF in P129 variety. H52 had the highest level of 4-4 dimethylsterols (355 mg/100g oil) at 28 DAF. The greatest amount of 4-monomethylsterols (35 mg/100g oil) was detected in H52 at 14 DAF. During linseed development, beta sitosterol (830 mg/100g oil) was the major 4-desmethylsterols, followed by campesterol (564 mg/100g oil) and stigmasterol (265 mg/100g oil). Some of these compounds followed nearly the same accumulation pattern during linseed maturation.

[Evolution of tocopherols in relation of unsaturated fatty acids during maturation of seeds of rapeseed (Brassica napus L.)]. / Evolution des tocophérols en relation avec les acides gras insaturés au cours de la maturation des graines de colza de printemps (Brassica napus L.).

The oil content increases during the maturation of seeds (rise of 30%), but decreases at the end of seed maturation. Differences between SDS-PAGE total protein profiles were shown. Polyunsaturated fatty acids contents increase during middle-maturation. Contents of alpha and gamma tocopherols increase with time. This increase is explained by the fact that tocopherols participate actively in the protection of membranes whose phospholipids consist of polyunsaturated fatty acids (PUFAs).

Evolution of phosphoenolpyruvate carboxylase activity and lipid content during seed maturation of two spring rapeseed cultivars (Brassica napus L.).

Phosphoenolpyruvate carboxylase (PEPc: EC 4.1.1.31) activity was monitored during seed maturation of two varieties (Hybridol and Pactol) of rapeseed (Brassica napus L.), widely cultivated in Tunisia. In the Hybridol variety, PEPc activity did not exceed 5 micromol h(-1) per gram of fresh weight (FW) during the first stages of maturation. It then highly increased to reach more than 30 micromol h(-1) g(-1)/FW. On the contrary, in the Pactol variety, the evolution of PEPc activity showed a classical curve, i.e. an increase during the most active phase of lipid accumulation in maturating seeds, followed by a rapid decrease until the end of seed maturation. In both varieties, the seed oil was characterised by a high content of oleic acid (C(18:1)), linoleic (C(18:2)) and linolenic acids (C(18:3)). Saturated fatty acids were also present, although decreasing with maturation course. The analysis of the triacylglycerols (TAG) showed that trioleoylglycerol (OOO) and dioleoyllinoleoylglycerol (OOL) were the major species (ca. 35% and ca. 25% of the total respectively). The evolution pattern of fatty acids and TAG contents was similar to that of PEPc activity. Taken together, our findings suggest that PEPc may be involved in fatty acid and triacylglycerol biosynthesis during seed maturation of both rapeseed varieties.