Design of three-component essential oil extract mixture from Cymbopogon flexuosus, Carum carvi, and Acorus calamus with enhanced antioxidant activity.

The development of novel antioxidant compounds with high efficacy and low toxicity is of utmost importance in the medicine and food industries. Moreover, with increasing concerns about the safety of synthetic components, scientists are beginning to search for natural sources of antioxidants, especially essential oils (EOs). The combination of EOs may produce a higher scavenging profile than a single oil due to better chemical diversity in the mixture. Therefore, this exploratory study aims to assess the antioxidant activity of three EOs extracted from Cymbopogon flexuosus, Carum carvi, and Acorus calamus in individual and combined forms using the augmented-simplex design methodology. The in vitro antioxidant assays were performed using DPPH and ABTS radical scavenging approaches. The results of the Chromatography Gas-Mass spectrometry (CG-MS) characterization showed that citral (29.62%) and niral (27.32%) are the main components for C. flexuosus, while D-carvone (62.09%) and D-limonene (29.58%) are the most dominant substances in C. carvi. By contrast, ß-asarone (69.11%) was identified as the principal component of A. calamus (30.2%). The individual EO exhibits variable scavenging activities against ABTS and DPPH radicals. These effects were enhanced through the mixture of the three EOs. The optimal antioxidant formulation consisted of 20% C. flexuosus, 53% C. carvi, and 27% A. calamus for DPPHIC50. Whereas 17% C. flexuosus, 43% C. carvi, and 40% A. calamus is the best combination leading to the highest scavenging activity against ABTS radical. These findings suggest a new research avenue for EOs combinations to be developed as novel natural formulations useful in food and biopharmaceutical products.

The structural characterization and bioactivity assessment of nonspecific lipid transfer protein 1 (nsLTP1) from caraway (Carum carvi) seeds.

BACKGROUND: Carum carvi (caraway) of the Apiaceae family has been used in many cultures as a cooking spice and part of the folk medicine. Previous reports primarily focus on the medicinal properties of caraway seed essential oil and the whole seeds extract. However, no effort has been made to study caraway proteins and their potential pharmacological properties, including nonspecific lipid transfer protein (nsLTP), necessitating further research. The current study aimed to characterize nonspecific lipid transfer protein 1 (nsLTP1) from caraway seed, determine its three-dimensional structure, and analyze protein-ligand complex interactions through docking studies. We also evaluated nsLTP1 in vitro cytotoxic effect and antioxidant capacity. Additionally, nsLTP1 thermal- and pH- stability were investigated. METHODS: Caraway nsLTP1 was purified using two-dimensional chromatography. The complete amino acid sequence of nsLTP1 was achieved by intact protein sequence for the first 20 residues and the overlapping digested peptides. The three-dimensional structure was predicted using MODELLER. Autodock Vina software was employed for docking fatty acids against caraway nsLTP1. Assessment of nsLTP1 cytotoxic activity was achieved by MTS assay, and the Trolox equivalent antioxidant capacity (TAC) was determined. Thermal and pH stability of the nsLTP1 was examined by circular dichroism (CD) spectroscopy. RESULTS: Caraway nsLTP1 is composed of 91 residues and weighs 9652 Da. The three-dimensional structure of caraway nsLTP1 sequence was constructed based on searching known structures in the PDB. We chose nsLTP of Solanum melongena (PDB ID: 5TVI) as the modeling template with the highest identity among all other homologous proteins. Docking linolenic acid with caraway protein showed a maximum binding score of -3.6 kcal/mol. A preliminary screening of caraway nsLTP1 suppressed the proliferation of human breast cancer cell lines MDA-MB-231 and MCF-7 in a dose­dependent manner with an IC50 value of 52.93 and 44.76 µM, respectively. Also, nsLTP1 (41.4 µM) showed TAC up to 750.4 µM Trolox equivalent. Assessment of nsLTP1 demonstrated high thermal/pH stability. CONCLUSION: To the best of our knowledge, this is the first study carried out on nsLTP1 from caraway seeds. We hereby report the sequence of nsLTP1 from caraway seeds and its possible interaction with respective fatty acids using in silico approach. Our data indicated that the protein had anticancer and antioxidant activities and was thermally stable.

Phytochemical constitutes and biological activities of essential oil extracted from irradiated caraway seeds (Carum carvi L.).

PURPOSE: Essential oils (EOs) obtained from spices, herbs, and medicinal plants are well known in traditional medicine and are an area of interest due to their various biological activities. Therefore, the present study investigates the chemical composition, phytochemical properties, as well as the biological activity of EOs, recovered from un-irradiated and irradiated (2.5, 5, and 10 kGy) caraway seeds. MATERIALS AND METHODS: Carum carvi L. seeds were irradiated with gamma irradiation at dose levels 2.5, 5, and 10 kGy, then EOs were recovered from all the samples. The chemical composition, phenols, and flavonoids content were evaluated. As well, antimicrobial and antitumor activities against the two cell lines [colorectal adenocarcinoma (Caco-2) and liver cancer (HepG-2)] were investigated. RESULTS: The results indicated the percentage of oil increased by radiation, especially a dose of 10 kGy, which gave the highest percentage (3.50%) compared to the control. Also, the Gas Chromatography/Mass Spectrometry (GC-MS) analysis revealed the presence of 26 compounds in the essential oil extracts. The main constituent of caraway seeds EOs was Carvone followed by Limonene. According to the results, there was an increase in the content of phenols and flavonoids by using gamma rays compared with control, the maximum increase was observed at dose level 10 kGy (13.70 and 7.38 mg/g oil, respectively) followed by 5 kGy (11.20 and 5.86 mg/g oil, respectively). The antioxidant properties of the caraway essential oils were increased by increasing the irradiation dose level (2.5-10 kGy) analyzed by DPPH radical and metal chelating activity. Caraway essential oils have an antimicrobial action against Gram-positive and Gram-negative bacteria as well as fungi. The antimicrobial activity was increased as the irradiation dose was raised and the10 kGy dose was more effective in suppressing the growth of bacteria and fungi. Additionally, the caraway essential oils have anticancer activity against the two cell lines studied; colorectal adenocarcinoma (Caco-2) and liver cancer cell line (HepG-2) as reduced the cell viability and density. CONCLUSION: The 10 kGy dose was more effective for oil yield, phenols, flavonoids, and antioxidant activity as well as antibacterial and antifungal activities. Furthermore, the caraway essential oils indicated anticancer activity against the two cell lines studied; colorectal adenocarcinoma (Caco-2) and liver cancer cell line (HepG-2) as reduced the cell viability and density. So caraway could be considered an important herb with multiple therapeutic uses.

Stimulation of Secondary Metabolites and γ-Terpinene Synthase by Silver Nanoparticles in Callus Cultures of Carum carvi.

Biotechnology and nanotechnology are important tools for understanding biochemical pathways. They can be used efficiently for stimulating and increasing the production of secondary metabolites in medicinal plants. The present study aimed to identify the γ-terpinene synthase gene (CcTPS2) as an effective contributor to the biosynthetic pathway of monoterpenes. The effects of silver nanoparticles (AgNPs; 50 and 100 mg l- 1) and time (24 and 48 h) were examined on secondary metabolites in cell suspension cultures of Carum carvi. This involved the identification, isolation, and sequencing of a partial sequence in the CcTPS2 gene of C. carvi. The genomic sequence of CcTPS2 comprised 292 bp which were organized into two exons (110 and 82 bp) and one intron (100 bp), while the cDNA was 192 bp. In the scale of nucleotides, the CcTPS2 gene showed 96% similarity with the TPS2 gene of Oliveria decumbens. We generated sequence data of the CcTPS2 gene for the first time in this species, thereby enabling further developments in understanding the molecular mechanisms responsible for terpene biosynthesis and other chemical derivatives in C. carvi. The results of GC/MS and GC/FID showed that AgNPs strongly affected the secondary metabolites in cell suspension cultures of C. carvi. According to the results, the AgNPs (50 mg l- 1) increased p-cymene and carvone contents in comparison with the control. The exposure of plants to 100 mg l- 1 AgNPs induced the production of thymol and carvacrol. The results of real-time PCR revealed that the exposure of plants to 100 mg l- 1 AgNPs caused a significant upregulation of CcTPS2 expression for 24 h. These cell suspension cultures were elicited by AgNPs, the application of which proved as an effective method to improve the production of secondary metabolites in vitro.

Low-dose gamma irradiation and pectin biodegradable nanocomposite coating containing curcumin nanoparticles and ajowan (Carum copticum) essential oil nanoemulsion for storage of chilled lamb loins.

The current investigation assessed the effect of pectin (PE) biodegradable nanocomposite coating containing curcumin nanoparticles (CNP) and ajowan (Carum copticum) essential oil nanoemulsion (ANE) combined with low-dose gamma irradiation on microbial, physiochemical, and sensorial qualities of lamb loins during refrigeration conditions. Active coating combined with gamma irradiation reduced the count number of mesophilic and psychrotrophic bacteria, lactic acid bacteria, Enterobacteriaceae; and minimized lipid and protein oxidation changes, total volatile basic nitrogen content, met-myoglobin formation, and color deterioration in the loin samples. The increased shelf-life of lamb loins up to 25 days compared with 5 days assigned for the control group can be associated with the application of ionizing radiation and edible PE coating containing CNP and ANE, which might be due to the synergistic or additive effects of treatments. Overall, as an effective preservation technique, a combination of PE + CNP + ANE and irradiation can be recommended for prolonging the shelf-life of lamb loins during refrigerated storage.

Characterization and Physical and Biological Properties of Tissue Conditioner Incorporated with Carum copticum L.

AIM: One of the main problems in dentistry is the injury caused by the long-term application of an ill-fitting denture. The existence of multiple microorganisms along with the susceptibility of the tissue conditioners to colonize them can lead to denture stomatitis. This study is aimed at developing a tissue conditioner incorporated with Carum copticum L. (C. copticum L.) for the effective treatment of these injuries. MATERIALS AND METHODS: The Carum copticum L. essential oil composition was determined by gas chromatography-mass (GC-mass) spectrometry. The antimicrobial activity of the essential oil against the standard strains of bacterial and fungal species was determined by broth microdilution methods as suggested by the Clinical and Laboratory Standards Institute (CLSI). The physical and chemical properties of the prepared tissue conditioner were investigated by viscoelasticity, FTIR assays, and the release study performed. Furthermore, the antibiofilm activity of the Carum copticum L. essential oil-loaded tissue conditioner was evaluated by using the XTT reduction assay and scanning electron microscopy (SEM). RESULTS: The main component of the essential oil is thymol, which possesses high antimicrobial activity. The broth microdilution assay showed that the essential oil has broad activity as the minimum inhibitory concentration was in the range of 32-128 µg mL-1. The viscoelasticity test showed that the essential oil significantly diminished the viscoelastic modulus on the first day. The FTIR test showed that Carum copticum L. essential oil was preserved as an independent component in the tissue conditioner. The release study showed that the essential oil was released in 3 days following a sustained release and with an ultimate cumulative release of 81%. Finally, the Carum copticum L. essential oil exhibited significant activity in the inhibition of microbial biofilm formation in a dose-dependent manner. Indeed, the lowest and highest amounts of biofilm formation on the tissue conditioner disks are exhibited in the Streptococcus salivarius and Candida albicans by up to 22.4% and 71.4% at the 64 µg mL-1 concentration of C. copticum L. with a statistically significant difference (P < 0.05). CONCLUSION: The obtained results showed that the Carum copticum L. essential oil-loaded tissue conditioner possessed suitable physical, biological, and release properties for use as a novel treatment for denture stomatitis.

Microencapsulated Caraway Essential Oil Affects Initial Growth of Maize Cultivars.

Caraway (Carum carvi L.) essential oil is a candidate for botanical herbicides. A hypothesis was formulated that the sand-applied maltodextrin-coated caraway oil (MCEO) does not affect the growth of maize (Zea mays L.). In the pot experiment, pre-emergence application of five doses of MCEO was tested on four maize cultivars up to the three-leaf growth stage. The morphological analyses were supported by the measurements of relative chlorophyll content (SPAD), two parameters of chlorophyll a fluorescence, e.g., Fv/Fm and Fv/F0, and fluorescence emission spectra. The analyzed MCEO contained 6.5% caraway EO with carvone and limonene as the main compounds, constituting 95% of the oil. The MCEO caused 7-day delays in maize emergence from the dose of 0.9 g per pot (equal to 96 g m-2). Maize development at the three-leaf growth stage, i.e., length of roots, length of leaves, and biomass of shoots and leaves, was significantly impaired already at the lowest dose of MCEO: 0.4 g per pot, equal to 44 g m-2. A significant drop of both chlorophyll a fluorescence parameters was noted, on average, from the dose of 0.7 g per pot, equal to 69 g m-2. Among the tested cultivars, cv. Rywal and Pomerania were less susceptible to the MCEO compared to the cv. Kurant and Podole. In summary, maize is susceptible to the pre-emergence, sand-applied MCEO from the dose of 44 g m-2.

Antispasmodic activity and mechanism of action of polyherbal formulation DCD-684 on rabbit jejunum.

Digas colic drops (DCD-684) is a polyherbal formulation containing decoctions of five medicinal plants namely Carum carvi L., Foeniculum vulgare Mill, Mentha arvensis L., Mentha piperita L. and Zingiber officinale Roscoe. These plants have been extensively used in traditional medicine for the treatment of various gastrointestinal diseases including abdominal colic. This study was conducted to determine the spasmolytic effect of DCD-684 (100% v/v) and its individual plant components on isolated rabbit jejunum (in vitro) and their possible mechanism of action. The effects were evaluated on spontaneous and pre-contracted tissues using KCl (80mM) and other contractile agonists including acetylcholine (0.3µM), carbamylcholine (0.3µM), serotonin (10 µM) and histamine (100µM) in the presence and absence of DCD-684. The various concentrations of DCD-684 (0.1-3% v/v) demonstrated spasmolytic effects on both spontaneous (IC50=0.75%) and KCl-induced contractions (IC50=1.6%), respectively. It also inhibited the contractions induced by acetylcholine (IC50=0.45%), carbamylcholine (IC50=0.95%), serotonin (IC50=0.95%) and histamine (IC50=0.87%). The DCD-684 exhibited synergistic effect due to its five plant components suggesting that spasmolytic cascade is probably governed by muscarinic and/or nicotinic receptors, serotonergic histaminergic, as well as calcium channel blocking mechanisms. Thereby, providing the pharmacological basis of its therapeutic use in the gastrointestinal motility disorders and related inflammatory ailments.

Bacterial and oxidative control of local butter with smart/active film based on pectin/nanoclay/Carum copticum essential oils/ß-carotene.

In this study, Pectin/Nanoclay (montmorillonite)/Carum copticum Essential oils/ß-Carotene (Pec/Clay/CCE/ßC) composite film was prepared. The effect of ß-Carotene (ßC) and Carum copticum Essential oils (CCE) concentration on the antioxidant activity and color/mechanical properties of Pec/Clay/CCE/ßC film was studied. The morphology and antibacterial activity of film were studied. The films containing maximum essential oil and ß-carotene showed the best antibacterial activity, antioxidant activity, flexibility and firmness. So the Pec/Clay/CCE0.5%/BC0.03% film as the optimum film was used for packaging of local butter. Oxidative stability, microbial count, and color properties (L*, a* and b*) of butter packaged with active film (Pec/Clay/CCE/ßC) were investigated. Results showed that Pec/Clay/CCE/ßC film was effective against Bacillus cereus (B. cereus) more than Escherichia coli (E. coli). According to the results active packaged butter had the highest oxidative stability, lowest microbial load, and the least color change during storage. In the packaging process, Pec/Clay/CCE/ßC film color was changed from orange to light yellow and this color change was used as a smart color indicator to detect the oxidation of butter and expiration time of butter.

Rapid Spectrophotometric Method for Assessing Hydroperoxide Formation from Terpenes in Essential Oils upon Oxidative Conditions.

Essential oils are widely used in the food and cosmetics industry as natural flavoring and fragrance substances. For this reason, a thorough quality control applying selected analytical methods is required. Oxidation along with hydroperoxide formation is an important drawback during production and storage of essential oils. Hydroperoxides constitute the main products formed upon photo-oxidation of essential oils. Due to hydroperoxide instability, gas chromatography (GC) and high-performance liquid chromatography (HPLC) analyses are required. According to the European Pharmacopoeia, titration is the official method for oxidation assessment. However, this analysis is time-consuming, and large sample quantities are required. Here, we present a simple and accurate spectrophotometric method for the detection of peroxide trace amounts in essential oils and terpenes. The principle is based on the formation of Wurster's red, which is enforced by the peroxide-driven oxidation of N,N-dimethyl-p-phenylenediamine dihydrochloride (DMPD). The method was validated using dibenzoyl peroxide (DBP) and cumene hydroperoxide (CHP). To demonstrate the suitability of the method for routine analysis, various oxidized terpenes and essential oils were chosen. Moreover, photo- and thermal oxidation experiments were compared and evaluated using gas chromatography/mass spectrometry (GC/MS) and a synthesized limonene-2-hydroperoxide (Lim-2-OOH) reference standard to gather detailed information on the structural changes of the respective terpenes.

Changes in beneficial bioactive compounds in eight traditional herbal liqueurs during a one-month maceration process.

BACKGROUND: Herbs are rich in various beneficial bioactive compounds and they can be used for many different purposes. One of the most common is maceration of herbs in alcohol. Different substances respond differently to extraction in prepared solutions. It is very important to enhance the highest proportion of beneficial ingredients during the maceration process in a herbal liqueur. A comparative analysis of numerous different phenolic compounds from eight of the most frequently used herbs for making aperitifs in Europe was performed. RESULTS: In the comparison among all studied herbs, the highest phenolic content was found with Artemisia absinthium (wormwood) (14.61 mg g-1 ) and Mentha piperita (peppermint) (13.89 mg g-1 ), while the lowest content was found with Centaurium erythraea (common centaury) (3.96 mg g-1 ). Salvianolic acid isomers and caffeic acid derivatives were the greatest contributors to the total phenolic content in lemon balm, wormwood, peppermint, fennel and sage. These compounds contain more hydroxyl groups and they were better extracted at the beginning of the maceration process. Caraway and common centaury contain more flavonols (quercetin and kaempferol derivatives), with higher chemical stability and fewer hydroxyl groups in their structure. CONCLUSION: The compositions of eight herbal liqueurs were highly related to the presence of different herbal ingredients and their solubility in the extract over time. Most flavonol derivatives were extracted over a longer time and the two liqueurs (common centaury and caraway) therefore had the highest phenolic contents after 3 weeks of maceration. © 2019 Society of Chemical Industry.

Bioactive extracts of Carum copticum and thymol inhibit biofilm development by multidrug-resistant extended spectrum ß-lactamase producing enteric bacteria.

The emergence and spread of multidrug-resistant (MDR) pathogenic bacteria is a clinical problem that requires novel anti-infective agents. Targeting pathogenic biofilms is considered a promising strategy to control bacterial infections. In this study, bioactive extracts of Carum copticum were investigated for their anti-biofilm efficacy against extended spectrum ß-lactamase (ESßL) producing MDR enteric bacteria. Thymol was also tested for its anti-biofilm properties, as gas chromatography-mass spectrometry revealed a high content (65.8%) of this phytochemical in the C. copticum methanolic extract. Biofilm inhibition was assessed in microtitre plates and further validated by light, electron and confocal laser microscopy. Sub-inhibitory concentrations of bioactive extracts of C. copticum and thymol significantly prevented biofilm development, ranging from 78.6 to 83.9% reductions. Microscopic analysis revealed that biofilms made by ESßL producing MDR enteric bacteria had a weakened structure, scattered microcolonies, and reduced cell density and thickness after exposure to the bioactive extracts and thymol.

Synthesis of Carum Carvi essential oil nanoemulsion, the cytotoxic effect, and expression of caspase 3 gene.

Scientists are attempting to find novel methods to overcome cancers. Nanoemulsion systems as the novel drug delivery tools have been widely used in cancer therapy. In this study, the Carum Carvi oil nanoemulsions (CCONE) were prepared and its cytotoxic activity was studied on human colon cancer HT-29 cells using MTT assay. Flow cytometry and Real-time qPCR were triggered to evaluate the nanoemulsions' apoptotic properties. The results showed a significant negative association between the HT-29 cancer cell viability and CCONE doses of treatments compared with Huvec normal cells (p value < 0.001). The IC50 values were estimated 12.5 µg/ml and 50 µg/ml for HT-29 and Huvec, respectively. Moreover, we observed that increasing concentrations of nanoemulsions significantly upregulate Caspase-3 gene expression. The results showed the CCONE is an efficient novel apoptosis inducer for human colon cancer cells without any undesirable side effects. However, further in vitro and in vivo researches are required. PRACTICAL APPLICATIONS: Cancer is a complex and usually untreatable disorder. Several types of cancer therapy strategies have been applied widely to overcome cancers. Chemotherapy has been used in various types of cancers. In most cases, not only it had not been effective on cancer cells but also been distractive within normal tissues. According to results, Carum Carvi essential oil nanoemulsions have apoptotic and cytotoxic effects on colon cancer cells (HT-29). When it comes to cancer of any kind, it's important to realize that no dietary supplement can fully treat, cure, or prevent cancer. However, there are some supplements that can potentially decrease the risk of cancer. Nanoemulsions present several advantages including the ability to incorporate hydrophilic, amphiphilic, and lipophilic excipient ingredients, high physical stability, and rapid gastrointestinal digestibility. The Carum Carvi essential oil nanoemulsion can also be applied as an effective food supplement due to its potent apoptotic activity.

para-Menthane as a Stable Terpene Derived from Orange By-Products as a Novel Solvent for Green Extraction and Solubilization of Natural Substances.

This study aims at investigating p-menthane, a novel bio-based solvent resulting from the hydrogenation of d-limonene, as a green alternative to n-hexane or toluene for the extraction and solubilization of natural substances. First, conductor-like combination of quantum chemistry (COSMO) coupled with statistical thermodynamics (RS) calculations show a comparable solubilization profile of p-menthane and n-hexane for carotene, volatile monoterpenes such as carvone and limonene, and model triglycerides. Other data obtained experimentally in solid/liquid extraction conditions further indicate that p-menthane showed similar performances to n-hexane for extracting carotenes from carrots, aromas from caraway seeds, and oils from rapeseeds, as these products showed a comparable composition. p-Menthane was also tested using common analytical extraction procedures such as Soxhlet for determination of oil content via multiple extraction stages, and Dean-Stark for determination of water content via azeotropic distillation. For both systems, yields were comparable, but for Dean-Stark, the distillation curve slope was higher when using p-menthane, and the time needed to attain 100% water recovery was 55% shorter than for toluene. Taken together, these results reveal the potential of p-menthane as a green replacer for petroleum-based solvents such as n-hexane or toluene.

Evaluation of Carum-loaded Niosomes on Breast Cancer Cells:Physicochemical Properties, In Vitro Cytotoxicity, Flow Cytometric, DNA Fragmentation and Cell Migration Assay.

Thymoquinone (TQ), a phytochemical compound found in Carum carvil seeds (C. carvil), has a lot of applications in medical especially cancer therapy. However, TQ has a hydrophobic nature, and because of that, its solubility, permeability and its bioavailability in biological mediums are poor. To diminish these drawbacks, we have designed a herbal carrier composed of Ergosterol (herbal lipid), Carum carvil extract (Carum) and nonionic surfactants for herbal cancer treatment. C. carvil was extracted and characterized by GC/Mass. Two different formulations containing TQ and Carum were encapsulated into niosomes (Nio/TQ and Nio/Carum, respectively) and their properties were compared together. Morphology, size, zeta potential, encapsulation efficiency (EE%), profile release rate, in vitro cytotoxicity, flow cytometric, DNA fragmentation and cell migration assay of formulations were evaluated. Results show that both loaded formulations have a spherical morphology, nanometric size and negative zeta potential. EE% of TQ and Carum loaded niosomes was about 92.32% ± 2.32 and 86.25% ± 1.85, respectively. Both loaded formulations provided a controlled release compared with free TQ. MTT assay showed that loaded niosomes have more anti-cancer activity compared with Free TQ and free Carum against MCF-7 cancer cell line and these results were confirmed by flow cytometric analysis. Cell cycle analysis showed G2/M arrest in TQ, Nio/TQ and Nio/Carum formulations. TQ, Nio/TQ and Nio/Carum decreased the migration of MCF7 cells remarkedly. These results show that the TQ and Carum loaded niosomes are novel carriers with high efficiency for encapsulation of low soluble phytochemicals and also would be favourable systems for breast cancer treatment.

Effect of Caraway, Fennel and Melissa addition on in vitro Rumen Fermentation and Gas Production.

BACKGROUND AND OBJECTIVE: Medicinal herbs and aromatic plants could be used to manipulate rumen fermentation. This study aimed to evaluate the effect of adding herbal and aromatic plants at 1, 3, 5, 7% of total ratio DM supplementation of the incubation media of an in vitro rumen model. MATERIAL AND METHODS: About 400±4 mg of feed sample (roughage and concentrate ratio of 45:55%) with each level, weighted into 125 mL glass bottles (6 bottles for each treatment), rumen fluid injected into these bottles and incubated at 39°C, after 24 h incubation digestibility of dry matter (IVDMD) and organic matter digestibility (IVOMD), total gas production (TG) and metabolic energy (ME) were studied. RESULTS: The differences among plants, added at different levels, were significant. Significant differences were also observed between highest level added compared with control in ammonia (NH3) and volatile fatty acids (VFA), IVDMD and IVOMD compared with control. Total gas (TG) was significantly higher at level 7%, especially with added Melissa compared with other plants. Metabolic energy (ME), was significantly higher in all treatments compared with control. CONCLUSION: It may be concluded that addition of different medicinal and aromatic herbal plants Caraway (Carum carvi), fennel (Foeniculum vulgare) and Melissa (Melissa officinali)), especially at highest levels tested has a great potential in manipulating rumen fermentation, which may be of benefit when applied in ruminant nutrition.

Study the antibacterial and antibiofilm activity of Carum copticum against antibiotic-resistant bacteria in planktonic and biofilm forms.

OBJECTIVES: Microbial biofilms has attracted interest in the recent years because they has become the most important cause of nosocomial infections. This study was aimed to examine the antibacterial activities of Carum copticum extracts on the development of microbial biofilms and planktonic form of six pathogenic bacteria. METHODS: Antimicrobial activity of the crude extracts against the planktonic form of six pathogenic bacteria: Bacillus cereus, Staphylococcus aureus, Pseudomonas aeruginosa, Escherichia coli, Acinetobacter baumannii and Klebsiella pneumonia was evaluated by using the disc diffusion method. Minimal Inhibitory Concentration (MIC) and Minimal Bactericidal Concentration (MBC) values was determined by macro-broth dilution technique. Anti-biofilm effects were assessed by microtiter plate method. The chemical composition of the herbal extract was identified by GC-MS. RESULTS: According to disc diffusion test (MIC and MBC) the ability of C. copticum extracts for inhibition of bacteria in planktonic form was confirmed. The best inhibitory effect of this plant on S. aureus and low inhibitory effect on A. baumannii in planktonic forms were observed. These extracts were efficient to inhibit biofilm structures and concentration of each extract has direct relation with inhibitory effect. The maximum and minimum inhibitory effects of C. copticum methanolic extract on biofilm formation were observed on A. baumannii (98%) and K. pneumoniae (19%) respectively. CONCLUSION: The GC-MS analysis revealed that five active compounds were present in the extract of this plant. Data obtained, suggested that the C. copticum extracts applied as antimicrobial agents against these pathogens particularly in biofilm making.

Anti-Oxidant and digestive enzymes inhibitory based anti diabetic activity of crude and fractions of Carum carvi L. extracts.

In the present study crude ethanolic extract and its various fractions (ethyl acetate, hexane and aqueous) of medicinal plant Carum carvi L. were examined for α-amylase and α-glucosidase inhibition using an-in vitro model. Both digestive enzymes were extracted from bovine and green gram. The crude extract and its fractions were also studied for their antioxidant potential by DPPH and Nitric oxide activity. The quantitative assessment of phenol and flavonoid contents was also estimated. The crude extract and its fractions exhibited high in-vitro enzyme inhibitory activity against α-amylase and α-glucosidase at different concentrations with IC50 ranging from 421.4±7.8 to 810±5.71and 72±8.81 to 307.0±11.42µg/mL of each extract respectively. The plant showed highest total phenolic contents ranging from 29.5±0.49 to 112.5±0.36mg/g Gallic acid of extract, while the total flavonoid contents were estimated from3.08±0.02-85.4± 0.12mg/g Quercetin. The antioxidant activities of the all extracts, measured in terms of IC50 values were in the range of 53.05±1.98 to 211.5±31.06µg/mL. Nitric oxide scavenging ability exhibited their IC50 values from 26.3±5.51 to 121.3±5.32µg/mL. Ethanolic crude extract showed excellent result among all these fractions. GCMS analysis of ethanolic extract of Carum carvi L indicated the presence of several phytochemicals such as monoterpenes, unsaturated fatty acids, furan derivatives, phenolic and flavonoid contents.

Chemical profiles and efficacy of essential oils obtained from three spices against Helicobacter pylori

Introduction: the medicinal herbs constitute one of the most preferred alternative solutions as antimicrobial agents due to their availability, biodegradability, fewer side effects. Unfortunately, rare are the studies focused on the efficacy of plant extracts against Helicobacter pylori to either confirm or refute their effectiveness. Objective: determinate the chemical profiles and evaluate the antibacterial activity against Helicobacter pylori of the essential oils obtained from Eugenia caryophyllata, Foeniculum vulgare and Carum carvi. Methods: the essential oils obtained from flower buds of Eugenia caryophyllata and seeds of Foeniculum vulgare and Carum carvi were extracted by hydrodistillation. The antibacterial potency against local clinical isolate of Helicobacter pylori was tested using agar disc diffusion and minimum inhibitory concentration techniques and the chemical composition was determined via gas chromatography-mass spectrometry. Results: the phytochemical analysis identified trans-anethole (43.01%), estragole (27.04%) and fenchone (06.63%) as main compounds in F. vulgare oil. Carvone and α -pinene were defined as major components of C. carvi oil with 63.92% and 8.43% respectively, whereas the abundant constituents of E. caryophyllata were eugenol (65.22%), eugenyl acetate (18.77%) and trans-caryophyllene (9.92%). The strongest anti H. pylori activity was exhibited by F. vulgare oil reaching similar action of Clarithromycin (p < 0.05) used as positive control. Conclusion: this is the first report showing the potency of essential oils from spices against H. pylori, the results indicated that these resources can constitute potential natural treatment.

Introducción: las hierbas medicinales constituyen una de las soluciones alternativas preferidas para desempeñar el papel de agentes antimicrobianos debido a su disponibilidad, biodegradabilidad y pocos efectos secundarios. Desafortunadamente, son raros los estudios centrados en la eficacia de los extractos de las plantas contra Helicobacter pylori que confirman o refutan su eficacia. Objetivo: evaluar la actividad antibacteriana contra Helicobacter pylori de los aceites esenciales de Eugenia caryophyllata, Foeniculum vulgare y Carum carvi. Métodos: los aceites esenciales obtenidos de los brotes de la flor del E. caryophyllatay, de semillas del F. vulgare y del C. carvi fueron extraídos por el hydrodistillation. La potencia antibacteriana contra el aislante clínico local Helicobacter pylori fue probada usando la difusión del disco del agar. Las técnicas de concentración inhibitoria mínima y la composición química fueron determinadas vía cromatografía del gas, espectrometría de masas. Resultados: los análisis fitoquímicas de los aceites esenciales identificado trans-anethole (43,01 %), estragole (27,04 %) y fenchone (6,63 %) como compuestos mayoritarios en F. vulgare. El aceite extraído del C. carvi reveló la presencia del carvone y α -pinene con los compuestos abundantes a 63,92 % y 8,43 % respectivamente, mientras que los componentes principales para E. caryophyllata eran el eugenol (65,22 %), eugenyl acetate (18,77 %) y trans-caryophyllene (9,92 %). De los aceites esenciales obtenidos, el de F. vulgare presentó mayor actividad anti-H. pylori, con la acción similar al del Clarithromycin (p < 0,05) usada como control positivo. Conclusión: los aceites esenciales de Eugenia caryophyllata, Foeniculum vulgare y Carum carvi pueden constituir un potencial tratamiento natural contra H. pylori.

Antibacterial activity of traditional spices against lower respiratory tract pathogens: combinatorial effects of Trachyspermum ammi essential oil with conventional antibiotics.

The aim of this study was to investigate the effects of ajowan essential oil (AjEO)/thymol and antibiotics combinations against three standard strains and six resistant clinical isolates of major respiratory bacteria (Pseudomonas aeruginosa, Staphylococcus aureus and Streptococcus pneumoniae). The broth microdilution method was conducted to determine the minimum inhibitory concentrations (MIC) of essential oil/thymol and antibiotics. The checkerboard method was used to investigate the interactions between the essential oil/thymol and antibiotics by means of the fractional inhibitory concentration index (FICI). The chemical composition of essential oil was also analysed by GC-MS and GC-FID. Thymol (50·75%), γ-terpinene (25·94%) and p-cymene (18·31%) were identified as major constituents of the oil. The most sensitive organisms to ajowan volatile oil were Strep. pneumoniae bacteria (MIC = 0·125-0·5 mg ml-1 ). Synergistic effects were observed with AjEO/thymol and amoxicillin combinations on methicillin-resistant Staph. aureus clinical isolates (FICI = 0·37-0·50) and with essential oil and ciprofloxacin combinations against P. aeruginosa ATCC 27853, Staph. aureus ATCC 25923 and penicillin (P)-resistant Strep. pneumoniae bacteria (FICI = 0·37-0·50). Combination of thymol and ciprofloxacin produces synergistic effects only against P. aeruginosa ATCC 27853 and P-resistant Strep. pneumoniae clinical isolate (FICI = 0·46-0·49).