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Purification and biological activity of acidic polysaccharide from leaves of Thymus vulgaris L.

Polysaccharides are involved in biological responses and can activate complement system, which plays an important role in primary host defense mechanisms. We investigated anticomplementary activities from spice plants and selected thyme (Thymus vulgaris L.) as a potent complementary activator. Acidic polysaccharide (TV-3-IIIA-IIa) purified from the hot-water extract of thyme leaves by DEAE-Toyopearl 650C, Butyl-Toyopearl 650M and Sepharose CL-6B column chromatography and preparative HPLC. The purified polysaccharide, TV-3-IIIA-IIa showed potent anticomplementary activity via classical and alternative pathway with the increase proportional to dosage. TV-3-IIIA-IIa seemed to be a homogenous polymer from the results of HPLC and its molecular mass was estimated as 180 kDa. TV-3-IIIA-IIa mainly consisted of galacturonic acid (44.8 mol%), glucuronic acid (16.7 mol%), arabinose (11.1 mol%), rhamnose (9.2 mol%), galactose (8.9 mol%) and small amounts of glucose, xylose, mannose and fucose. By methylation analysis and reactivity to beta-glucosyl Yariv reagent, TV-3-IIIA-IIa was assumed to contain small amounts of type II arabinogalactan and large amounts of pectin-like polysaccharides in the structure. Based upon these results, TV-3-IIIA-IIa was suggested to be a complement activator.

Animals↗

Purification and characterization of anti-complementary polysaccharide from leaves of Thymus vulgaris L.

For the purification of the anti-complementary polysaccharide from Thymus vulgaris L., the hot-water extract of thyme leaves was successively fractionated by ethanol precipitation and ultra-filtration. The retentate with 300-kDa membrane cartridge showed a potent anti-complementary activity. It was further purified by open column chromatographies on DEAE-Toyopearl 650C, Butyl-Toyopearl 650M and Sephadex G-100, obtaining TV3-IIA-I, the purified anti-complementary polysaccharide. The anti-complementary polysaccharide exhibited the anti-complementary activity via both classical and alternative pathways. Based upon the methylation analysis and the reaction with beta-glucosyl Yariv reagent, the anti-complementary polysaccharide from thyme might contain an arabinogalactan moiety, at least in part.

Chromatography, Ion Exchange↗

Adult repellency and larvicidal activity of five plant essential oils against mosquitoes.

The larvicidal activity and repellency of 5 plant essential oils--thyme oil, catnip oil, amyris oil, eucalyptus oil, and cinnamon oil--were tested against 3 mosquito species: Aedes albopictus, Ae. aegypti, and Culex pipiens pallens. Larvicidal activity of these essentials oils was evaluated in the laboratory against 4th instars of each of the 3 mosquito species, and amyris oil demonstrated the greatest inhibitory effect with LC50 values in 24 h of 58 microg/ml (LC90 = 72 microg/ml) for Ae. aegypti, 78 microg/ml (LC90 = 130 microg/ml) for Ae. albopictus, and 77 microg/ml (LC90 = 123 microg/ml) for Cx. p. pallens. The topical repellency of these selected essential oils and deet against laboratory-reared female blood-starved Ae. albopictus was examined. Catnip oil seemed to be the most effective and provided 6-h protection at both concentrations tested (23 and 468 microg/ cm2). Thyme oil had the highest effectiveness in repelling this species, but the repellency duration was only 2 h. The applications using these natural product essential oils in mosquito control are discussed.

Aedes↗

Estrogen and progestin bioactivity of foods, herbs, and spices.

In this study we report on the content and bioactivity of plant (phyto) estrogens and progestins in various foods, herbs, and spices, before and after human consumption. Over 150 herbs traditionally used by herbalists for treating a variety of health problems were extracted and tested for their relative capacity to compete with estradiol and progesterone binding to intracellular receptors for progesterone (PR) and estradiol (ER) in intact human breast cancer cells. The six highest ER-binding herbs that are commonly consumed were soy, licorice, red clover, thyme, tumeric, hops, and verbena. The six highest PR-binding herbs and spices commonly consumed were oregano, verbena, tumeric, thyme, red clover and damiana. Some of the herbs and spices found to contain high phytoestrogens and phytoprogestins were further tested for bioactivity based on their ability to regulate cell growth rate in ER (+) and ER (-) breast cancer cell lines and to induce or inhibit the synthesis of alkaline phosphatase, an end product of progesterone action, in PR (+) cells. In general, we found that ER-binding herbal extracts were agonists, much like estradiol, whereas PR-binding extracts, were neutral or antagonists. The bioavailability of phytoestrogens and phytoprogestins in vivo were studied by quantitating the ER-binding and PR-binding capacity of saliva following consumption of soy milk, exogenous progesterone, medroxyprogesterone acetate, or wild mexican yam products containing diosgenin. Soy milk caused a dramatic increase in saliva ER-binding components without a concomitant rise in estradiol. Consumption of PR-binding herbs increased the progestin activity of saliva, but there were marked differences in bioactivity. In summary, we have demonstrated that many of the commonly consumed foods, herbs, and spices contain phytoestrogens and phytoprogestins that act as agonists and antagonists in vivo.

Alkaline Phosphatase↗

Effects of herbal essential oils used to extend the shelf life of freshwater-reared Asian sea bass fish (Lates calcarifer).

Sensory and microbiological characteristics of Asian sea bass (Lates calcarifer) fish raised in a freshwater pond were evaluated during cold storage for 33 days. Whole fish (averaging 400 g each) were stored in a cold storage room at 0 to 2 degrees C. Essential oils of herbs--thyme (Thymus vulgaris) and oregano (Origanum vulgare)--added at 0.05% (vol/vol) were used as preservatives. On the basis of the sensory test results as well as the bacteriological tests, the addition of oregano and thyme essential oils can considerably slow the process of spoilage. The fish treated with these oils were still fit for human consumption after 33 days of storage. The results obtained through sensory tests are corroborated to a great extent by the chemical (hypoxanthine) tests and to a lesser extent by the physical (Cosmos units) tests. The initial total bacteriological counts were 1.7 x 10(3) CFU/cm2 on the fish surface and <10(2) CFU/g in the fish flesh, and in the control treatment (without preservatives), these counts rose continuously, reaching around 10(7) CFU/cm2 on the fish surface and 10(3) CFU/g in the flesh after 33 days of storage at 0 to 2 degrees C. The use of herbal essential oils as preservatives, on the other hand, resulted in a maximal count of 10(5) CFU/cm2 on the fish surface, while the bacterial count in the flesh remained <10(2) CFU/g by the end of the 33-day storage period at 0 to 2 degrees C.

Animals↗

Antibacterial efficiency of Finnish spice essential oils against pathogenic and spoilage bacteria.

The antibacterial properties of 13 essential oils, derived from spices grown in Finland, were examined with an agar diffusion method against 12 bacterial strains. The organisms tested included both spoilage and pathogenic bacteria. The gram-positive bacteria appeared to be more sensitive than the gram-negative organisms, Clostridium botulinum and Clostridium perfringens being the most sensitive. Oregano, savory, and thyme showed the broadest antibacterial activity by distinctly inhibiting the growth of all the organisms tested. By gas chromatography-mass spectrometry analysis, differences were noted in the composition of oregano and thyme oils in comparison to previous reports.

Anti-Bacterial Agents↗

Antibacterial activities of naturally occurring compounds against antibiotic-resistant Bacillus cereus vegetative cells and spores, Escherichia coli, and Staphylococcus aureus.

After demonstrating the lack of effectiveness of standard antibiotics against the acquired antibiotic resistance of Bacillus cereus (NCTC 10989), Escherichia coli (NCTC 1186), and Staphylococcus aureus (ATCC 12715), we showed that the following natural substances were antibacterial against these resistant pathogens: cinnamon oil, oregano oil, thyme oil, carvacrol, (S)-perillaldehyde, 3,4-dihydroxybenzoic acid (beta-resorcylic acid), and 3,4-dihydroxyphenethylamine (dopamine). Exposure of the three pathogens to a dilution series of the test compounds showed that oregano oil was the most active substance. The oils and pure compounds exhibited exceptional activity against B. cereus vegetative cells, with oregano oil being active at nanogram per milliliter levels. In contrast, activities against B. cereus spores were very low. Activities of the test compounds were in the following approximate order: oregano oil > thyme oil approximately carvacrol > cinnamon oil > perillaldehyde > dopamine > beta-resorcylic acid. The order of susceptibilities of the pathogens to inactivation was as follows: B. cereus (vegetative) >> S. aureus approximately E. coli >> B. cereus (spores). Some of the test substances may be effective against antibiotic-resistant bacteria in foods and feeds.

Anti-Bacterial Agents↗

Collaborative study of a method for the extraction of light filth from whole, cracked, or flaked and ground spices.

An improved method has been developed for the extraction of light filth from whole, cracked, or flaked spices (basil, bay leaves, clery leaves, chervil, chives, dill weed, mint flakes, parsley, rosemary, sage, tarragon, thyme, and vegetable flakes) and from ground spices (cloves, cumin, marjoram, mustard seed, oregano, sage, and thyme). The method involves a chloroform or isopropanol defatting, followed by a direct flotation from 40% isopropanol with Tween 80-EDTA (1+1) and mineral oil-heptane (85+15). Collaborative results show that the proposed method is more rapid to perform and yields better filth recoveries than the official first action methods for ground spices, 44.116, and whole, cracked, or flaked spices, 44.129. The method has been adopted as official first action.

Animals↗

Investigation of in vitro antimicrobial activity of honey.

Apitherapy or therapy with bee products as honey is an old tradition. In this study, the antimicrobial activity of 84 honey samples were investigated against 8 bacteria potential pathogens and 2 fungi. Honey samples were obtained from 10 different floral sources. The findings indicate that honey samples with thyme, thyme + acorn, pine + carob, pine + carob + anis and pine + chestnut are more effective than the other honey of floral sources. The results of the survey show that most of honey samples at 50% (w/v) can completely inhibit the growth of all of the tested bacteria. The fungi were less sensitive than the bacteria to the antimicrobial activity of honey. The sensitivity of microorganisms to sugar solutions with similar sugar contents as some honey samples which have high antimicrobial activity were also assayed. This sugar solutions inhibited a narrow range of bacteria and did not inhibit any of the fungi tested. In conclusion, these results encourage further studies to investigate possible benefits of the use of honey among therapies in the treatment of bacterial infections.

Anti-Bacterial Agents↗

Inhibitory activity of spices and essential oils on psychrotrophic bacteria.

This study was designed to evaluate "in vitro" the inhibitory effects of spices and essential oils on the growth of psycrotrophic food-borne bacteria: Aeromonas hydrophila, Listeria monocytogenes and Yersinia enterocolitica. The sensitivity to nine spices and their oils (chilli, cinnamon, cloves, ginger, nutmeg, oregano, rosemary, sage, thyme) was studied. Antibacterial activity was evaluated on liquid and solid medium. Spices: 1% concentration of each spice was added separately to Triptic Soy Broth and then inoculated to contain 10(8)/ml organism and held to 4 degrees C for 7 days. Populations of test organism were determined on Triptic Soy Agar. Oils: Inhibition of growth was tested by using the paper disc agar diffusion method (at 35, 20 and 4 degrees C) and measuring their inhibition zone. MIC was determined by the broth microdilution method. Some culinary spices produce antibacterial activity: inhibition of growth ranged from complete (cinnamon and cloves against A. hydrophila) to no inhibition. Antibacterial inhibition zone ranged from 8 mm to 45 mm: thyme essential oil showed the greatest inhibition against A. hydrophila.

Aeromonas hydrophila↗

Immunologic reactivity to work-related airborne allergens in people occupationally exposed to dust from herbs.

A group of 150 people occupationally exposed to dust from herbs were examined. The examined group consisted of 47 thyme farmers, 32 chamomile farmers, 31 sage farmers and 40 workers of herbs processing industry. As a reference group, 50 urban dwellers, not exposed to any kind of organic dust, were examined. Skin prick tests and precipitin tests were conducted with, respectively, 4 and 11 microbial antigens associated with organic dust. Both skin and precipitin tests were also conducted with herbal extracts of chamomile and sage. Precipitin tests were carried out with sera not concentrated and sera 3-fold concentrated. Tests for inhibition of leukocyte migration (MIF) were also conducted with 4 microbial antigens. People occupationally exposed to dust from herbs showed a higher frequency of positive skin reactions to microbial antigens compared to the reference group. The results of precipitin test also revealed greater reactivity to the environmental microbial antigens in the examined group, compared to the reference group. The highest frequency of positive results was noted with the antigen of Pantoea agglomerans (30.6 % with sera not concentrated and 48.3 % with sera 3-fold concentrated) - the difference compared to the reference group (12.0 %) was highly significant (p < 0.01). The frequencies of positive results of MIF test in the examined group were high with all antigens tested: Arthrobacter globiformis (12.6 %), Pantoea agglomerans (11.1 %), Saccharopolyspora rectivirgula (17.0 %), Aspergillus fumigatus (13.3 %), and, compared to the reference group with no positive result for any antigen, all the differences were significant (p < 0.05). In conclusion, the frequency of positive allergological reactions to airborne microorganisms was high in people occupationally exposed to dust from herbs and suggests a potential role of microbial allergens in the pathogenesis of work-related health disorders among herb workers. The risk of sensitization seems to be greatest among thyme farmers, who showed the highest positive response. The results confirmed the particular allergenic importance of Gram-negative bacterium Pantoea agglomerans.

Adolescent↗

[Development of throat clearing herbal teas].

Medicinal herbs in tea for throat clearing are used from ancient times. Taking into consideration the bronchial mucus secretion stimulating and antispasmodic, antimicrobial, antiphlogistic and stimulating effect on the ciliated epithelium two new formulations of throat clearing herbal tea were originated. The first formulation consists of liquorice roots, sweet fennel and thyme, and the second one consists of pine gemmae, thyme and elder flowers. The methods for identification and assay of the active substances of the components were adapted. The purity of the mixtures was regulated by the limitation of the loss on drying, total ash, microbial contamination, contamination with radionuclides, heavy metals, pesticides and foreign matter. Expiry date of both throat clearing herbal teas was confirmed to be 2 years.

Drug Stability↗

Inhibitory effects of spices and herbs towards Shigella sonnei and S. flexneri.

The use of spices and herbs, their essential oils or their active compounds as means of control of pathogens constitutes an alternative to chemical additives In the present study the antibacterial activities of cloves, thyme, oregano, rosemary and basil on Shigella have been established. Although in a model system in agar media addition of 1% basil could contribute to the 'hurdle' principle and delay outgrowth of Shigella sp. this was not confirmed in real food conditions: the presence of 1% basil did not affect growth of Shigella in potato puree at 22 degrees C or survival at 7 degrees C in spaghetti sauce. Thyme and essential oils and thymol and carvacrol showed inhibition of Shigella sp. in the agar well diffusion method (MIC 0.1-1.0%) and they have potential to be used as a desinfectant in the washing water e.g. in the process line of minimal processed vegetables. However, more studies combining sensoric properties with microbial analysis are needed to investigated the possible use of these compounds.

Colony Count, Microbial↗

Respiratory effects of exposure to dust from herbs.

A group of 150 people occupationally exposed to dust from herbs were examined. As a reference group, 50 urban dwellers, not exposed to any kind of organic dust were examined. People were subjected to routine physical examination and to lung function examinations with the LUNGTEST 500 spirometer (MES, Kraków, Poland). The spirometric values of the forced expiratory volume in one second (FEV (1)), vital capacity (VC) and FEV(1)/VC were recorded before and after work. Physical chest examination revealed pathological crepitations in 10 people (6.7 %). The mean baseline spirometric values in the study and reference groups did not show significant differences compared to the normal values. In the herb workers exposed to organic dust the post-shift decrease of all analysed spirometric values was noted. The post-shift decrease of some spirometric values (VC, VC % of normal values) was highly significant (p < 0.01). There was evidenced of a significant positive correlation between the age of examined people and decrease of VC and FEV(1) values. In 12 exposed workers the decrease of FEV(1) or FEV(1) % of normal values higher than 15 % was noted. 50 % of these workers cultivated thyme (Thymus vulgaris L.). This may suggest that dust from herbs, especially thyme dust, may cause acute airway obstruction. In the group showing significant decrease of FEV(1)/FEV(1) % of normal values ( > 15 %) the frequency of reported respiratory work-related symptoms (83.3 %) was higher than in the rest of exposed group (61.5 %). In conclusion, occupational exposure to dust from herbs may cause harmful effects on the respiratory system among herb processing workers. This indicates the need for use of prophylactic measures in this professional group, the more so as number of people occupationally exposed to dust from herbs is growing.

Adolescent↗

The use of Thymi Herba as feed additive (0.1%, 0.5%, 1.0%) in weanling piglets with assessment of the shedding of haemolysing E. coli and the detection of thymol in the blood plasma.

Thymi Herba (Thymus vulgaris, rubbed) was given as feed additive to weanling piglets. 3 concentrations (0.1%, 0.5%, 1%) were tested against a control group. Rectal swabs were collected weekly and were sent in for bacterial testing. The shedding of haemolysing E. coli was evaluated. There was no significant difference in the shedding of haemolysing E. coli between the 4 groups. Neither was there a difference in the distribution of the various serotypes. Thymol was detected in the blood plasma in all thyme groups. The increase in thymol level with greater amounts of thyme herb was significant. Three days after withdrawal of the feed additive no thymol levels were detected. The study results did not reveal any effects on haemolysing E. coli in the gut.

Animals↗

[Study on anti-tumor effect of medicinal fungi Phellinus igniarius extracts].

OBJECTIVE: To study the inhibiting effect of medicinal fungi Phellinus igniarius extracts on S180 tumor and the immunoregulation effect on the S180-induced tumor mice. METHOD: S180 mice were orally given 100, 200, 400 mg x kg(-1) dosage of P. igniarius extracts, then the inhibition grow effect, spleen index, and thyme index were measured. RESULT: Medicinal fungi P. igniarius extracts can increase the spleen index and thyme index and the inhibiting tumor rate was 31.88%, 46.25%, 53.13%, respectively. Also, medicinal fungi P. igniarius extracts can prolong life in mice. CONCLUSION: The medicinal fungi P. igniarius extracts show obviously anti-tumor effect and immunoregulation effect.

Animals↗

Potential mold growth, aflatoxin production, and antimycotic activity of selected natural spices and herbs.

Ground spices and herbs are evaluated as substrates for mycelial growth, sporulation, and aflatoxin production. Three toxigenic strains of Aspergilli, A. flavus ATCC 15548, A. flavus NRRL 3251, and A. parasiticus NRRL 2999, were cultured on moist, commercially packaged herbs and spices. All substrates used were ground and included thyme, celery seed, oregano, cinnamon, ginger, caraway seed, clove, mustard, sesame seed, and rosemary leaves. Following inoculation of the natural materials in sterile bottles containing sterile water, the cultures were incubated 30 days at 23 +/- 4 degrees C. Not all strains of Aspergilli grew, sporulated, or produced toxins. There were definite strain differences and definite substrate differences for the variables evaluated. Sesame seed produced toxins B1, G1, and G2, with a mean of 167 ppm for 3 strains. A. flavus ATCC 15548 was the greatest overall toxin producer followed by A. parasiticus NRRL 2999 and A. flavus NRRL 3251. Ginger and rosemary leaves were also substantial producer-substrates. Mustard, caraway seed, and celery seed were judged as intermediate-producing substrates. Absolute antimycotic substrates were cinnamon and clove. Antiaflatoxigenic substrates were thyme and oregano. Mustard also may be antimycotic. Aflatoxins B1 and G1 were the more commonly found toxins.

Aflatoxins↗

Randomized trial of aromatherapy. Successful treatment for alopecia areata.

OBJECTIVE: To investigate the efficacy of aromatherapy in the treatment of patients with alopecia areata. DESIGN: A randomized, double-blind, controlled trial of 7 months' duration, with follow-up at 3 and 7 months. SETTING: Dermatology outpatient department. PARTICIPANTS: Eighty-six patients diagnosed as having alopecia areata. INTERVENTION: Eighty-six patients were randomized into 2 groups. The active group massaged essential oils (thyme, rosemary, lavender, and cedarwood) in a mixture of carrier oils (jojoba and grapeseed) into their scalp daily. The control group used only carrier oils for their massage, also daily. MAIN OUTCOME MEASURES: Treatment success was evaluated on sequential photographs by 2 dermatologists (I.C.H. and A.D.O.) independently. Similarly, the degree of improvement was measured by 2 methods: a 6-point scale and computerized analysis of traced areas of alopecia. RESULTS: Nineteen (44%) of 43 patients in the active group showed improvement compared with 6 (15%) of 41 patients in the control group (P = .008). An alopecia scale was applied by blinded observers on sequential photographs and was shown to be reproducible with good interobserver agreement (kappa = 0.84). The degree of improvement on photographic assessment was significant (P = .05). Demographic analysis showed that the 2 groups were well matched for prognostic factors. CONCLUSIONS: The results show aromatherapy to be a safe and effective treatment for alopecia areata. Treatment with these essential oils was significantly more effective than treatment with the carrier oil alone (P = .008 for the primary outcome measure). We also successfully applied an evidence-based method to an alternative therapy.

Adult↗