Promising Antileishmanial Activity of Micromeria nervosa Essential Oil: In Vitro and In Silico Studies.

The present study aimed to evaluate the leishmanicidal potential of the essential oil (EO) of Micromeria (M.) nervosa and to investigate its molecular mechanism of action by qPCR. Furthermore, in silicointeraction study of the major M. nervosa EO compounds with the enzyme cytochrome P450 sterol 14α-demethylase (CYP51) was also performed. M. nervosa EO was analyzed by gas chromatography-mass spectrometry (GC-MS). Results showed that α-pinene (26.44%), t-cadinol (26.27%), caryophyllene Oxide (7.73 ± 1.04%), and α-Cadinene (3.79 ± 0.12%) are the major compounds of M. nervosa EO. However, limited antioxidant activity was observed, as this EO was ineffective in neutralizing DPPH free radicals and in inhibiting ß-carotene bleaching. Interestingly, it displayed effective leishmanicidal potential against promastigote (IC50 of 6.79 and 5.25 µg/mL) and amastigote (IC50 of 8.04 and 7.32 µg/mL) forms of leishmania (L.) infantum and L. major, respectively. Molecular mechanism investigation showed that M. nervosa EO displayed potent inhibition on the thiol regulatory pathway. Furthermore, a docking study of the main components of the EO with cytochrome P450 sterol 14α-demethylase (CYP51) enzyme revealed that t-cadinol exhibited the best binding energy values (-7.5 kcal/mol), followed by α-cadinene (-7.3 kcal/mol) and caryophyllene oxide (-7 kcal/mol). These values were notably higher than that of the conventional drug fluconazole showing weaker binding energy (-6.9 kcal/mol). These results suggest that M. nervosa EO could serve as a potent and promising candidate for the development of alternative antileishmanial agent in the treatment of leishmaniasis.

Grape seed extract prevents chlorpyrifos-induced toxicity in rat liver through the modulation of phase I detoxification pathway.

Chlorpyrifos (CPF) poisoning is a public health problem for which there is not currently any effective prophylaxis. In this study, we investigated the protective effect of grape seed extract (GSE) against CPF-induced hepatotoxicity. Rats were daily treated either with CPF (2 mg/kg) or CPF and GSE (20 mg/kg) for 1 week, sacrificed, and their livers dissected for biochemical, molecular, and histopathological analyses. CPF generated liver dysfunction by altering carbohydrate, lipid, amino acid, ammonia and urea metabolism, and provoked mitochondrial impairment through disturbing tricarboxylic acid (TCA) cycle, oxidative phosphorylation (OXPHOS), and mitochondrial viability. CPF also induced cholinergic excitotoxicity along with oxidative stress and histopathological alterations. Interestingly, treatment with GSE prevented all the detrimental effects of CPF through the regulation of cytochrome P450 (CYP450) gene expression. Molecular docking analysis indicated that GSE-containing polyphenols acted as epigenetic modulators through inhibiting DNA (cytosine-5)-methyltransferase 1 (DNMT1), thus favoring the CYP2C6 detoxification pathway. Thereby, GSE might be a promising strategy in the protection of the liver against CPF toxicity.

Laboratory assessment of the acaricidal, repellent and anti-cholinesterase effects of Melaleuca alternifolia and Chamaemelum nobile essential oils against Hyalomma scupense ticks.

In cattle, Hyalomma scupense serves as an important vector of several pathogens resulting in diseases, subsequently affecting the agricultural field as well as the economy. Resistance to chemical acaricides has become widespread affirming the need for new drugs to tick control. The goal of this study was to investigate the acaricidal, repellent activities as well as the putative mode of action of two essential oils (EOs) from Melaleuca alternifolia (Tea tree) and Chamaemelum nobile (Roman chamomile) on Hyalomma scupense. The chemical composition of EOs was also evaluated. Different concentrations of EOs were tested in vitro for their acaricidal property on adults and larvae of H. scupense using adult immersion test (AIT) and larval packet test (LPT). Additionally, using Ellman's spectrophotometric method, the anticholinesterase (AChE) inhibition activity of M. alternifolia and C. nobile EOs was assessed in order to understand their putative mode of action. The main compounds of C. nobile were α-Bisabolene (22.20%) and (E)-ß-Famesene (20.41%). The major components in the analyzed M. alternifolia were Terpinen-4-ol (36.32%) and γ-Terpinene (13.69%). Adulticidal and larvicidal assays demonstrated a promising efficacy of the essential oils against tick H. scupense. The lethal concentration (LC50) values obtained for M. alternifolia and C. nobile oils were 0.84 and 0.96 mg/mL in the AIT and 0.37 and 0.48 mg/mL in the LPT, respectively. Regarding repellent activity, M. alternifolia achieved 100% repellency at the concentration of 1 mg/mL while C. nobile showed 95.98% repellency activity at concentration of 4 mg/mL. Also, M. alternifolia and C. nobile EOs displayed potent AChE inhibition with IC50 value of 91.27 and 100.12 µg/mL, respectively. In the present study, M. alternifolia and, to a lesser degree, C. nobile EOs were found to be effective in vitro acaricides, repellents and acetylcholinesterase inhibitor against H. scupense ticks. These plants may represent an economical and sustainable alternative to toxic synthetic acaricides in the management of ectoparasites of veterinary importance.

Screening of antimicrobial and insecticidal properties of essential oils extracted from three Tunisian aromatic and medicinal plants.

The essential oils (EOs) of Coriandrum sativum, Carum carvi and Artemisia herba-alba have been screened for their insecticidal activity against Tribolium castaneum, Sitophilus oryzae and Lasioderma serricorne and for their antimicrobial activity against Gram (+), Gram (-) bacteria and against yeast strain. Artemisia herba-alba EO showed good insecticidal activity with 24-h against L. serricorne (LC50 = 29.7) and against T. castaneum (66.1 µg/mL) and antibacterial potentials against Staphyloccocus aureus (MIC = 0.125 mg/mL). The antimicrobial activities of C. carvi EO, rich in D-carvone (72.4%) and D-limonene (23.8%) were particularly remarkable, with LC50 of 27.9 µg/mL against L. serricorne. Similarly, coriander EO, with linalool (64.6%) as the major compound was selected for its antimicrobial activities (Candida albicans, MIC = 1 mg/mL). These findings demonstrated that the tested EOs could be used as a natural source of insecticidal and antimicrobial activity, which may have a variety of uses in the food and pharmaceutical sectors.

Synergistic combination of Cinnamomum verum and Syzygium aromaticum treatment for cutaneous leishmaniasis and investigation of their molecular mechanism of action.

Combination therapy at appropriately suitable doses presents a promising alternative to monotherapeutic drugs. In this study, Cinnamomum verum and Syzygium aromaticum essential oils and their major compounds have exhibited substantial leishmaniacidal potential against both promastigote and amastigote forms of Leishmania (L.) major. However, they displayed high cytotoxicity against Raw264.7 macrophage cells. Interestingly, when combined with each other or with amphotericin B, they demonstrated a synergistic effect (FIC<0.5) with low cytotoxicity. These combinations are able to modulate the production of nitric oxide (NO) by macrophages. Notably, the combination of S. aromaticum Essential oil with amphotericin B stimulates macrophage cells by increasing NO production to eliminate leishmanial parasites. Furthermore, investigation of the molecular mechanism of action of these synergistic combinations reveals potent inhibition of the sterol pathway through the inhibition of the CYP51 gene expression. The findings suggest that combination therapy may offer significant therapeutic benefits in both food and pharmaceutical fields.

Pistacia lentiscus essential oil and its pure active components as acaricides to control Dermanyssus gallinae (Acari: Mesostigmata).

The goal of this work was to assess the in vitro acaricidal effects of Pistacia lentiscus essential oil and its pure active components against red poultry mites Dermanyssus gallinae as an alternative to chemical acaricide (formamidines). Essential oil was obtained using hydrodistillation and then analyzed using GC-MS. The following major components were identified: α-pinene (20.58 %), D-Limonene (18.16 %), ß-Myrcène (15.06 %), 4-Terpineol (7.68 %), caryophyllene (5.45 %) and γ-terpinene (5.21 %). The toxicity of essential oil and its six monoterpenes were tested at concentrations ranging from 0.43 to 3.50 mg/cm2, using contact and spraying bioassays. Toxicity was expressed as a lethal concentration (LC50 or LC90). The experiment results revealed that activity was method and concentration dependent, and the spraying method was more effective than the contact bioassay for acaricidal activity. In this bioassay, the highest mortality was observed with 4-Terpineol. The LC50 was estimated to be 0.184 mg/cm2 for this substance, followed by α-pinene, the LC50 of which was estimated to be 0.203 mg/cm2. Caryophyllene and γ-terpinene were found less effective in controlling D. gallinae. P. lentiscus oil and its major compounds were also evaluated for anti-acetylcholinesterase (AChE) effects; 4-Terpineol was found to be the most effective AChE inhibitor with IC50 values reaching 18.73 ± 2.83 µg/mL. This framework pointed out the importance of the traditional use of P. lentiscus as an ecofriendly alternative against ectoparasite of veterinary importance; D. gallinae. In vivo trials should also be conducted to assure the safe use of essential oils or individual compounds and to achieve efficient acaricidal property.

Toxicity, repellency, and anti-cholinesterase activities of bioactive molecules from clove buds Syzygium aromaticum L. as an ecological alternative in the search for control Hyalomma scupense (Acari: Ixodidae).

Introduction: The goal of the current study is to evaluate the acaricidal and repellent properties of the ethanolic extract, essential oil, and primary component eugenol from Syzygium aromaticum against Hyalomma scupense cattle ticks. Their potential mechanisms of action were also examined, using an in vitro assay. Methods: Clove essential oil was extracted using hydrodistillation technique. Gas chromatography-mass spectrometry (GC-MS) was performed to identify the chemical composition of clove. To evaluate the adulticidal, ovicidal, larvicidal and repellent proprieties of clove essential oil, eugenol and ethanolic extract on H. scupense, in vitro assays were performed using the adult immersion test (AIT), the ovicidal test, the larval packet test (LPT), the filter paper test and anti-acetylcholinesterase (AChE) activity. Results: After treatment, eugenol, the primary phytoconstituent of clove oil, which accounts for 97.66% of the whole oil, had 99.22% acaricide activity and inhibited egg hatching at a concentration of 10 mg/mL. Eugenol and clove essential oil showed potent adulticidal effect at high concentrations (10 mg/mL), achieving 100 and 93.76% mortality, respectively. The ethanolic extract exhibited moderate activity. At high concentration, the larvicidal activity of S. aromaticum oil, eugenol, and ethanolic extract were 100, 100, and 77.18%, respectively. In filter paper experiments, when tested at the concentration 5 mg/mL; eugenol showed the longest repellent effect up to 6 h. We also found that eugenol was the most active AChE inhibitor (IC50 = 0.178 mg/mL). Nevertheless, additional investigations are required to confirm the accurate mechanism and the relevance of clove in practical application. Conclusion: Overall, our research indicated that, because its effectiveness as acaricide, S. aromaticum essential oil and its phytoconstituent eugenol may offer an alternative source for the control of H. scupense cattle ticks.

Anti-Candida and Anti-Leishmanial Activities of Encapsulated Cinnamomum verum Essential Oil in Chitosan Nanoparticles.

Nanoencapsulation is widely considered as a highly effective strategy to enhance essential oils' (EO) stability by protecting them from oxidative deterioration and evaporation. The present study aims to optimize and characterize an efficient technique for encapsulating Cinnamomum (C.) verum essential oil into chitosan nanoparticles using response surface methodology (RSM). Moreover, the optimized C. verum EO nanoparticle was investigated for its antibacterial (against Gram-positive and Gram-negative bacteria), antifungal (against Candida albicans), and antiparasitic activity (against Leishmania parasites). Five parameters were investigated using a Plackett-Burman and Box-Behnken statistical design: the chitosan molecular weight, TPP concentration, C. verum EO/chitosan ratio, mixing method, and the duration of the reaction. Encapsulation efficiency and anti-candida activity were considered as responses. The antibacterial, anticandidal, and anti-leishmanial activities were also assessed using a standard micro-broth dilution assay and the cytotoxicity assay was assessed against the macrophage cell line RAW 264.7. The optimized nanoparticles were characterized using Fourier transform infrared spectroscopy, Zeta potential, and scanning electron microscopy. The study results indicated that under optimal conditions, the nanoencapsulation of C. verum EO into chitosan nanoparticles resulted in an encapsulation efficiency of 92.58%, with a regular distribution, a nanoparticle size of 480 ± 14.55 nm, and a favorable Zeta potential of 35.64 ± 1.37 mV. The optimized C. verum EO/chitosan nanoparticles showed strong antifungal activity against C. albicans pathogens (CMI = 125 µg mL-1), notable antibacterial activity against both Gram-positive and Gram-negative bacteria (ranging from 125 to 250 µg mL-1), high leishmanicidal potential against the promastigotes form of L. tropica and L. major (IC50 = 10.47 and 15.09 µg mL-1, respectively), and a four-fold cytotoxicity reduction compared to non-encapsulated essential oil. These results suggest that C. verum EO-loaded chitosan nanoparticles could be a promising delivery system for the treatment of cutaneous Candida albicans infections.

Nano-Emulsification of Cinnamon and Curcuma Essential Oils for the Quality Improvement of Minced Meat Beef.

This work aims to evaluate cinnamon and curcuma essential oils as natural preservatives in minced beef meat. Essential oil chemical compositions and antibacterial activities were studied, and their encapsulation was optimized into nano-emulsions based on droplet size and distribution assessments. Selected formulas were further explored for their physical stabilities and antibacterial activities. Then, their effects on minced beef meat preservation were evaluated. Results showed significant differences in the chemical compositions and the efficiency of the tested essential oils, with cinnamon having a significant antibacterial efficacy. Formulation results showed that cinnamon nanoemulsion, encapsulated by 7.5% Tween 80, possessed an 89 nm droplet size, while the droplet diameter of curcuma nanoemulsion, encapsulated by 5% Tween 80, was 151 nm. Antimicrobial results depicted a significantly higher activity in nanoemulsions as compared to essential oils. For instance, the inhibition diameter of cinnamon essential oils against S. aureus was equal to 35 mm, while that of its nanoemulsion reached 40 mm. The meat preservation results showed that both bulk and nanoencapsulated essential oils significantly inhibited bacterial growth, as well as the formation of methemoglobin and lipid oxidation in meat. Thus, this work draws attention to the enhanced preservation effects of essential oils on the processing of minced beef meat as well as the great potential of nanoemulsions as carriers for essential oils in food industry applications.

Acaricidal and anthelmintic efficacy of Ocimum basilicum essential oil and its major constituents estragole and linalool, with insights on acetylcholinesterase inhibition.

The present study evaluated the acaricidal and anthelmintic action of Ocimum basilicum essential oil and its main components against ticks and helminth parasites as well as to relate these activities to acetylcholinesterase inhibition. The in vitro acaricidal activity against Hyalomma scupense was evaluated by Adult Immersion Test (AIT) and Larval Packet Test (LPT), while the in vivo nematocidal potential was assessed in laboratory mice infected with Heligmosomoides polygyrus using fecal egg count reduction (FECR) and total worm count reduction (TWCR). Chemical analyzes were performed by gas chromatography coupled to mass spectrometry (GC-MS). Estragole (80.87%) and linalool (16.12%) were the major compounds detected in O. basilicum essential oil. In the AIT assay for H. scupense tick, LC50 of estragole, O. basilicum oil and linalool were 0.73, 0.81 and 0.97 mg/mL, respectively. In LPT, estragole, linalool and essential oil showed LC50 of 0.22, 1.11 and 1.19 mg/mL, respectively. Against He. polygyrus, the highest activity was observed with estragole administered at 100 mg/kg body weight (bwt), which resulted in a FECR of 90.86% and a TWCR of 82.91%. The O. basilicum essential oil, estragole and linalool inhibited the enzyme acetylcholinesterase (AChE) extracted from both parasites species. Estragole was found the most active AChE inhibitor with IC50 of 0.176 mg/mL for H. scupense and IC50 of 0.138 mg/mL for He. polygyrus larvae. The results of the present study pointed out the importance of the traditional use of O. basilicum as an eco-friendly alternative against endo and ectoparasites. In vivo trials should also be conducted to confirm the above-mentioned activities and to assure the safe use of natural plants.

Efficacy of synergistic activity of seed oils from Carthamus tinctorius (Safflower) and Nasturtium officinale (Watercress) on the lethality of the cattle tick Hyalomma scupense (Acari: Ixodidae).

Background: Ticks and tick-borne diseases are a severe economic and public-health problem for cattle producers. The emergence of acaricide resistance to synthetic chemical treatments has prompted interest in developing alternative tick control methods. Aim: The main objective of the current research was to identify the chemical structure of Carthamus tinctorius and Nasturtium officinale seed oils and to assess their anti-tick properties against Hyalomma scupense ticks both alone and in combination (1:1). Methods: Analytical methods were used to analyze the chemical components. For in vitro assays, adults of H. scupense were immersed in C. tinctorius and N. officinale seed oils at 100, 125, 200, and 300 mg/ml concentrations; for 5 minutes. Larvae of H. scupense were dipped in 25, 50, 100, 125, 200, and 300 mg/ml doses of seed oils; the mortality percentage was determined after 24 hours. Results: The seed oil safflower was mainly composed of linoleic acid (84.48%), followed by palmitic acid (6.54%) and stearic acid (3.77%). Meanwhile, watercress seed oil was mainly composed of linolenic acid (50.78%), gondoic acid (13.57%), linoleic acid (10.58%), palmitic acid (8.02%), and erucidic acid (6.62%). The Adults Immersion Test showed the sensitivity of ticks to C. tinctorius and N. officinale seed oil: C. tinctorius seed oil caused (95%) mortality of H. scupense at 300 mg/ml, while N. officinale seed oil induced (88.68%) mortality at the same concentration. At a 200 mg/ml concentration, C. tinctorius and N. officinale oil combined caused 100% mortality. Tested oils showed larvicidal efficacy. LC50 values for C. tinctorius and N. officinale seed oils were 84.16 and 61.78 mg/ml, respectively, in 24 hours. LC50 value of oils association (50% C. tinctorius: 50% N. officinale) was 47.96 mg/ml.The mixture of seed oils from two plants tested against H. scupense larvae and adult females at a 1:1 ratio showed synergistic interaction. Conclusion: Seed oils tested alone, and the mixture could be used as an alternative solution in the fight against ticks.

Phytochemistry, anti-tick, repellency and anti-cholinesterase activities of Cupressus sempervirens L. and Mentha pulegium L. combinations against Hyalomma scupense (Acari: Ixodidae).

The adulticidal, larvicidal, and repellent effects of Cupressus sempervirens, Mentha pulegium, and their combination (C. sempervirens and M. pulegium) against Hyalomma scupense were investigated, as well as their putative mode of action. The evaluation on H. scupense was performed with the adult immersion test at concentrations ranging from 2.5-20 mg/mL and the larval packet test from 0.625 to 20 mg/mlL. The same concentrations were used for combined oils. Essential oils were obtained using hydrodistillation and then analyzed using GC-MS. The following major components were identified: α-pinene (56.89 %) and δ-3-Carene (15.61 %) for C. sempervirens, while pulegone (70.50 %) and cis-Menthone (18.26 %) for M. pulegium. At 20 mg/mL concentration, C. sempervirens had (94.40 %) acaricide action and inhibited the egg from hatching, however at 10 mg/mL, it had 100 % larvicidal activity. Adulticidal and larvicidal activities of M. pulegium oil reached to 87.19 % and 81.02 % respectively, at 20 mg/mL concentration. The highest acaricidal property against H. scupense was exhibited by combination oils with minimum LC50 values of 1.76 mg/mL on adultes and 0.60 mg/mL on larvae. The repellent activities were 100, 95.80 and 100 %, at the 20 mg/mL concentration used in 6 h, for C. sempervirens, M. pulegium and mixture oil respectively. Our findings revealed that the mixture of C. sempervirens and M. pulegium, at the ratio 1:1 (v: v) of each oil, showed high synergistic effect (SF > 1) and the use of combination oil (C. sempervirens 50 %: M. pulegium 50 %) is more toxic against H.scupense. Furthermore, the volatile oils examined revealed significant inhibition of H.scupense acetylcholinesterase activity AChE. Based on the findings of the present framework, these essential oils have the potential to be applied as substitute for synthetic acaricides in the management of H.scupense bovine ticks.

Antibacterial and antibiofilm activity of Peganum harmala seed extract against multidrug-resistant Pseudomonas aeruginosa pathogenic isolates and molecular mechanism of action.

Biofilm formation of the opportunistic pathogen Pseudomonas (P). aeruginosa is one of the major global challenges to control nosocomial infections due to their high resistance to antimicrobials and host defense mechanisms. The present study aimed to assess the antibacterial and the antibiofilm activities of Peganum (P). harmala seed extract against multidrug-resistant P. aeruginosa isolates. Chemical identification of the active compound and determination of its molecular mechanism of action were also investigated. Results showed that P. harmala n-butanol "n-BuOH" extract exhibited antibacterial activity against multidrug-resistant P. aeruginosa isolates. This extract was even more active than conventional antibiotics cefazolin and vaamox when tested against three P. aeruginosa multidrug-resistant isolates. In addition, P. harmala n-BuOH extract exhibited potent bactericidal activity against PAO1 strain at MIC value corresponding to 500 µg/mL and attained 100% killing effect at 24 h of incubation. Furthermore, P. harmala n-BuOH extract showed an antibiofilm activity against P. aeruginosa PAO1 and exhibited 80.43% inhibition at sub-inhibitory concentration. The extract also eradicated 83.99% of the biofilm-forming bacteria. The active compound was identified by gas chromatography-mass spectrometry as an indole alkaloid harmaline. Transcriptomic analysis showed complete inhibition of the biofilm-related gene pilA when PAO1 cells were treated with harmaline. Our results revealed that P. harmala seed extract and its active compound harmaline could be considered as a candidate for a new treatment of multidrug-resistant P. aeruginosa pathogens-associated biofilm infections.

In vitro acaricidal activity of essential oil and crude extracts of Laurus nobilis, (Lauraceae) grown in Tunisia, against arthropod ectoparasites of livestock and poultry: Hyalomma scupense and Dermanyssus gallinae.

The current study assayed the toxicity of Laurus nobilis essential oil and crude extracts obtained using solvents of increasing polarity (cyclohexane, acetone and ethanol), on two ectoparasites of veterinary importance, i.e., Hyalomma scupense and Dermanyssus gallinae. The major components detected in bay laurel essential oil were dominated by 1.8-cineole (46.56 %), α-terpinenyl acetate (13.99 %), sabinene (7.69), α-pinene (5.75), linalool (5.50), methyleugenol (5.36 %) and ß-pinene (3.97). The highest total phenolic and flavonoids contents were present in the ethalonic extract of L. nobilis leaves at an amount of 152.88 mg gallic acid equivalents per gram of dry weight (GAE/g DW) and 21.77 mg quercetin equivalent per gram of dry weight (QE/g DW), respectively. In vitro acaricidal effects of essensial oil and crude extract of L. nobilis against H. scupense were ascertained by adult immersion test of engorged females (AIT) and larval packet test (LPT) compared with a reference drug amitraz. The essential oil exhibited strong acaricidal activity against tick engorged female and inhibition of hatching eggs. After 24 h of exposure, at the highest tested concentration (100 mg/mL) essential oil induced 90.67 % mortality of H. scupense larvae (LC50 = 10.69 mg/mL). Otherwise, essential oil exhibited high acaricidal activity compared to extracts, and among the extract, the ethanolic extract revealed the highest acaricidal efficacy (81.27 % female mortality). Results from mite contact toxicity showed that essential oil and extracts from L. nobilis were toxic to D. gallinae. Bay essential oil was both more toxic to mites, and faster in exerting this toxicity than other tested crude extracts. L. nobilis essential oil concentration leaded to enhance mortality of D. gallinae reaching the highest (100 %) mortality at 12 h with a concentration of 320 mg/mL. While, ethanolic extract acheived this rate after 24 h of exposure at same concentration. Cyclohexanic extract showed weak acaricidal activity.

Physiological responses and expression of sugar associated genes in faba bean (Vicia faba L.) exposed to osmotic stress.

Faba bean (Vicia faba L.) is the major food legume crop in Tunisia. However, its growth and yield is strongly affected by water-limited environments. In this study, osmotic stress exhibited a negative effect on Bachar and Badii cultivar. Nevertheless, the deteriorating effects of osmotic stress were relatively low on studied parameters of Bachar due to its better efficiency to reduce oxidative damage by increasing enzymatic activities such as catalase (CAT), superoxide dismutase (SOD) and ascorbate peroxidase (APX), accumulation of total chlorophyll (Chlt), soluble sugars and leaf relative water content (RWC). GC-MS analysis determined a total of 11 soluble carbohydrates induced by osmotic stress and differentially accumulated in the both cultivars. Bachar showed elevated levels of mannose, glucose, galactose, ribose, rhamnose and myo-inositol which might help to maintain osmotic adjustment, membranes and proteins protection from the damaging effect of reactive oxygen species. Sugar metabolism related genes (VfNINV3, VfPHS2, VfFRK4, VfHXK1, VfGPI1, VfSTP1.1, VfpGlcT1.1, VfSTP5.1, VfpGlcT1.2, VfSWEET2.1, VfVINV2, VfSUS1, VfPGM1, VfSUT1.1, VfGPT1, VfSPS1, VfSPP1, VfPHS1, VfSUT4.1 and VfTMT1.1) were differentially expressed in both cultivars demonstrating their important roles in sugar accumulation. Most of these genes were upregulated in the leaves of Bachar under moderate and severe stress, which could lead to increase glycolysis and tricarboxylic acid cycle in order to accelerate energy production, necessary to increase osmotic regulation and consequently enhancing the osmotic stress tolerance in that cultivar. Overall, sugars accumulation ability can be used as a useful indicator for the osmotic stress tolerant potential in faba bean breeding programs. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at (10.1007/s12298-021-00935-1).

Natural colourants analysis and biological activities. Association to molecular markers to explore the biodiversity of Opuntia species.

INTRODUCTION: Synthetic colourants are subject to various allergies. As a result, the use of plant-based additives has proved promising. OBJECTIVE: It is in this perspective that our objective was undertaken to investigate natural colourants from edible cactus. Opuntia species populations were single-sequence repeat (SSR) fingerprinted, the analysis of betalains and phenolic compounds and the evaluation of their biological potentials were also characterised. RESULTS: Photometric quantification of betalains and phenols showed an interspecific variation across Opuntia species. Opuntia ficus indica fruits showed the highest betalain [betaxanthins; 843.67 and betacyanins; 1400 mg/100 g dry weight (DW)] and polyphenol contents. Reversed-phase high-performance liquid chromatography (RP-HPLC) analysis showed that the variation of individual phenols profile was influenced by interspecific and genetic factors. Isorhamnetin-O-(di-deoxyhexosyl-hexoside) was the major compound and its content varied according to Opuntia species, while catechol was the predominant phenolic compound in O. humifusa with 1.88 µg/g DW. Concerning cactus species, Opuntia colourants exhibited a potent antiradical activity [half maximal inhibitory concentration (IC50 ) up to 1 µg/mL]. Opuntia species were effective against Gram-positive and Gram-negative bacterial strains [inhibition zone (IZ) up to 27 mm]. A high genetic diversity within Opuntia genotypes based on SSR markers was revealed. UPGMA (unweighted pair group method with arithmetic mean) dendrogram and PCoA (principal coordinate analysis) based on natural pigments and antimicrobial profiles indicated significant variation. The correlation approach proved the presence of a probably metabolic relationship between genetic markers, pigments and their biological activities. CONCLUSION: A possible association between molecular approach and metabolic profile analysis of Opuntia allows tracing the relationship among species for its genetic conservation.

Biological control of the soft rot bacterium Pectobacterium carotovorum by Bacillus amyloliquefaciens strain Ar10 producing glycolipid-like compounds.

Four hundred and fifty bacteria were evaluated for antagonistic activity against bacterial soft rot of potato caused by Pectobacterium carotovorum sp strain II16. A strain Ar10 exhibiting potent antagonist activity has been identified as Bacillus amyloliquefaciens on the basis of biochemical and molecular characterization. Cell free supernatant showed a broad spectrum of antibacterial activity against human and phytopathogenic bacteria in the range of 10-60 AU/mL. Incubation of P. carotovorum cells with increasing concentrations of the antibacterial compound showed a killing rate of 94.8 and 96% at MIC and 2xMIC respectively. In addition, the antibacterial agent did not exert haemolytic activity at the active concentration and has been preliminary characterized by TLC and GC-MS as a glycolipid compound. Treatment of potato tubers with strain Ar10 for 72 h significantly reduced the severity of disease symptoms (100 and 85.05% reduction of necrosis deep / area and weight loss respectively). The same levels in disease symptoms severity was also recorded following treatment of potato tubers with cell free supernatant for 1 h. Data suggest that protection against potato soft rot disease may be related to glycolipid production by strain Ar10. The present study affords new alternatives for anti-Pectobacterium carotovorum bioactive compounds against the soft rot disease of potato.