Sigla storax (Liquidambar orientalis) mitigates in vitro methane production without disturbances in rumen microbiota and nutrient fermentation in comparison to monensin.

AIM: The aim of this study was to investigate the in vitro dose-dependent effects of sigla storax (Styrax liquidus) on rumen microbiota and rumen microbial fermentation in comparison to monensin as a positive control. METHODS AND RESULTS: This study was carried out using a rumen simulation model (Rusitec). Treatments consisted of no additive (control), 10 mg l-1 of monensin sodium salt, 100 mg l-1 (Low-Sigla), and 500 mg l-1 (High-Sigla) of sigla storax (n = 6/treatment). In addition to rumen fermentation characteristics, rumen microbial composition was investigated using 16S rRNA sequencing. The methane variables and the acetate to propionate ratio decreased in the both High-Sigla and monensin groups (P < 0.05). High-Sigla had no effect on ammonia, total SCFA and nutrition degradation, while monensin decreased these parameters (P < 0.05). Unlike monensin, the sigla storax treatments did not affect the alpha or beta diversity indexes of the microbiota. The relative abundance of Methanomethylophilaceae and Ruminococcaceae decreased with High-Sigla and monensin (P < 0.05), and Atopobiaceae and Eggerthellaceae decreased with the both doses of sigla storax as well as monensin treatments (P < 0.05). Syntrophococcus, DNF00809, and Kandleria were among the genera that most decreased with High-Sigla and monensin (Q < 0.07) and were strongly positively correlated with methane production (r = 0.52-0.56). CONCLUSIONS: The high dose of sigla storax (500 mg l-1) decreased methane in the rumen ecosystem without adverse effects on nutrient degradation and SCFA production, and without dramatically impacting the microbial composition. Sigla storax might be a novel feed additive to mitigate methane in cattle.

Seasonal Variability in Pyrrolizidine Alkaloids in Jacobaea alpina from the Trentino-Alto Adige Region (Northern Italy).

In recent years, increased attention has been given to plants containing toxic pyrrolizidine alkaloids (PAs). Jacobaea alpina (syn. Senecio cordatus) is a tall forb growing on mountain pastures and meadows containing such alkaloids and therefore, the plant is considered as a noxious weed in these environments. The repartition of toxic macrocyclic PAs in the plant and their evolution during the vegetation period has been studied in two populations. Eight PAs were found where senciphylline and senecionine accounted in most samples for more than 85 % of total alkaloids. Leaves in April and stems in May started with high PA concentrations (19-22 mg/g dry matter), then alkaloid levels declined. This decrease was more rapid in stems than in leaves. Depending on the population, fully developed inflorescences in June and July PA contents were higher or lower than in the respective leaves. Later, also in the inflorescences PA concentration decreased. Combined with growth data total alkaloid content in the whole plant as mg/plant was highest in midsummer and declined afterwards. Finally, new emerging leaves in September had high PA levels, which declined markedly towards the end of the season in November. In sum, over a large period PA concentration appeared to be high enough to present a health risk for grazing animals.

Diversity of Secondary Metabolites in Roots from Conium maculatum L.

BACKGROUND: Conium maculatum is known as highly toxic plant, due to piperidine alkaloids present in the aerial parts. In a first attempt, in various tap root samples, however, alkaloids could not be detected. The present study describes active compounds in the tap roots from 16 populations harvested at maturity. The compounds were extracted with dichloromethane from root pieces of single plants and analyzed by gas chromatography-mass spectrometry. Ten bioactive compounds were evaluated: five furocoumarins, two prenylated coumarins, two aliphatic C17-polyacetylenes and the phenylpropanoid elemicin. A high variability could be observed, the highest concentrations were measured for falcarindiol, xanthotoxin and isopimpinellin, the lowest for elemicin. In sum C. maculatum roots contained comparable amounts of compounds that are characteristic for Apiaceae, and also occur in vegetables as carrots, parsnip, parsley or celeriac.

Chemodiversity of Essential Oils in Seseli libanotis (L.) W.D.J.Koch (Apiaceae) in Central Europe.

Seseli libanotis is an aromatic umbelliferous plant distributed sporadically on dry grassland edges in Europe and Western Asia. The essential oil composition in the different plant parts was studied from plants collected on nine sites in Austria and one site in Alto Adige, Italy. Monoterpenes such as α-pinene, sabinene and ß-myrcene and the sesquiterpene germacrene D were present in all essential oils from the aerial parts. Inflorescences and fruits had the highest essential oil contents. These essential oils from four sites were rich in acorenone B, while from other three sites they had carotol as a major component. Osthole as furocoumarin occurred in some oils. The root essential oils were dominated by α-pinene. The essential oil variability has been studied by principal component analysis (PCA) and discriminant analysis (DA) with plant parts or sampling site as a priori groups. PCA could well separate inflorescence and fruit essential oil samples from leaf and stem essential oil samples. DA differentiated well between the plant parts and most of the sampling sites.

Thymol in the intestinal tract of broiler chickens after sustained administration of thyme essential oil in feed.

Plant compounds occurring in phytogenic feed additives are involved in different pharmacological activities in the animal organism. Since the digestive tract acts as a first line of defence against foreign compounds, it is necessary to outline its response to dietary supplementation with bioactive plant components. Little information is available on the bioactivity of thymol as the main bioactive compound of Thymus vulgaris L. essential oil (TEO). The main objective of the present study was to provide a detailed view of the concentrations of thymol in plasma and the content of individual intestinal segments (duodenum, jejunum, ileum, caecum and colon) of broiler chickens after 4 weeks of dietary supplementation with different TEO concentrations. 32 one-day old Ross 308 hybrid broilers were randomly divided into four dietary treatment groups (0.00%, 0.01%, 0.05%, 0.1% w/w of TEO in the diet). Thymol concentrations in the duodenal chyme presented around 7% on average from the thymol amount administered in the feed. A significantly increased thymol amount was observed after 0.1% TEO addition to the diet compared with 0.01% TEO enrichment in the duodenal wall and gut content of jejunum, ileum, caecum and colon (p < 0.05). Thymol levels in the colon were significantly higher than in the ileum and about 1.7 times higher on average than those in the caecum. Significant coefficient of correlation was observed between thymol concentrations in plasma and feed, gut content of all intestinal segments as well as duodenal wall. Our results point to intensive thymol absorption in the initial sections of the digestive tract. In the current study, the role of intestine in biotransformation of thymol was observed, and it would be desirable to investigate whether thymol itself or thymol metabolites are responsible for beneficial effects in intestine.

Biodiversity within Melissa officinalis: Variability of Bioactive Compounds in a Cultivated Collection.

Phytochemical characters were evaluated in a five-year-old lemon balm collection consisting of 15 and 13 subspecies officinalis and altissima accessions, respectively. Stems were lower in essential oil than leaves. First cut leaves (June) gave more oil than those of the second cut (August). Subspecies officinalis plants had leaf oils rich in geranial, neral and citronellal in various proportions in the first cut. However, in the second cut the oils from all accessions appeared very similar with 80-90% geranial plus neral. Leaf oils of subsp. altissima contained sesquiterpenes (ß-caryophyllene, caryophyllene oxide, germacrene D) and also further monoterpenes in the second cut. Leaves had higher rosmarinic acid (RA) contents than stems. More RA was in subsp. officinalis than subsp. altissima leaves. First cut leaves were richer in RA than those from second cut. Total phenolics and antioxidant parameters showed that lemon balm is a valuable source of plant antioxidants.

Chemical composition and antimicrobial activity of garlic essential oils evaluated in organic solvent, emulsifying, and self-microemulsifying water based delivery systems.

The chemical composition of garlic essential oils (GEOs) extracted from two different cultivars has been characterized using GC-MS analysis. GEO that was extracted from the white-skin cultivar (WGO) had a lower percentage of the major constituents diallyl trisulfide and diallyl disulfide (45.76 and 15.63%) than purple-skin cultivar (PGO) which contained higher percentages (58.53 and 22.38%) of the same components, respectively. Evaluation of the antimicrobial activity of WGO and PGO delivered in organic solvent (isopropanol) showed dose-dependent antimicrobial activity against the tested pathogenic bacteria and fungi, especially with WGO. On the other hand, formulation of both GEOs in water-based emulsions totally suppressed the antimicrobial activity of GEO. Re-formulation of GEOs in water-based microemulsion (particle size 10.1nm) showed better antimicrobial activity than emulsions at the same concentration of GEOs. This study can assist in designing the proper water-based delivery system of GEO for application in food preservation.

Rumen microbial abundance and fermentation profile during severe subacute ruminal acidosis and its modulation by plant derived alkaloids in vitro.

Rumen microbiota have important metabolic functions for the host animal. This study aimed at characterizing changes in rumen microbial abundances and fermentation profiles using a severe subacute ruminal acidosis (SARA) in vitro model, and to evaluate a potential modulatory role of plant derived alkaloids (PDA), containing quaternary benzophenanthridine and protopine alkaloids, of which sanguinarine and chelerythrine were the major bioactive compounds. Induction of severe SARA strongly affected the rumen microbial composition and fermentation variables without suppressing the abundance of total bacteria. Protozoa and fungi were more sensitive to the low ruminal pH condition than bacteria. Induction of severe SARA clearly depressed degradation of fiber (P < 0.001), which came along with a decreased relative abundance of fibrolytic Ruminococcus albus and Fibrobacter succinogenes (P < 0.001). Under severe SARA conditions, the genus Prevotella, Lactobacillus group, Megasphaera elsdenii, and Entodinium spp. (P < 0.001) were more abundant, whereas Ruminobacter amylophilus was less abundant. SARA largely suppressed methane formation (-70%, P < 0.001), although total methanogenic 16S rRNA gene abundance was not affected. According to principal component analysis, Methanobrevibacter spp. correlated to methane concentration. Addition of PDA modulated ruminal fermentation under normal conditions such as enhanced (P < 0.05) concentration of total SCFA, propionate and valerate, and increased (P < 0.05) degradation of crude protein compared with the unsupplemented control diet. Our results indicate strong shifts in the microbial community during severe SARA compared to normal conditions. Supplementation of PDA positively modulates ruminal fermentation under normal ruminal pH conditions.

Effect of Thyme Essential Oil Supplementation on Thymol Content in Blood Plasma, Liver, Kidney and Muscle in Broiler Chickens.

The absorption and metabolism of phytogenic feed additives in poultry is studied related to the metabolism and deposition of their main compounds in tissues intended for food production. Fifty-six non-sexed Ross 308 broilers were allocated to seven dietary treatments and fed a diet containing graded levels of thyme (Thymus vulgaris L.) essential oil (EO) (0, 0.01, 0.02, 0.03, 0.04, 0.05, 0.1%, w/w). Thymol concentration was measured in plasma, liver, kidney and breast muscle tissue using solid phase micro-extraction followed by gas chromatography/mass spectrometry. We found the highest concentrations of thymol in kidney and plasma, and the lowest in breast muscle and liver. Thymol content in plasma and kidney significantly increased when 0.05 and 0.1%, w/w, EO and in liver and breast muscle only when 0.1%, w/w, EO was added to the diet (p<0.05). Our results indicate intensive metabolism of thymol in liver and its accumulation in kidney tissue. We confirm low deposition of thymol in the muscle tissue. It is necessary to.-keep in mind the selection of a sufficient concentration of EO in the feed additive for animals without the risk of thymol residues in edible tissues.

Pyrrolizidine Alkaloids in Adenostyles alliariae and A. glabra from the Austrian Alps.

The alkaloid content of Adenostyles alliariae and A. glabra (Asteraceae) has been evaluated. Both species contain toxic macrocyclic unsaturated pyrrolizidine alkaloids with seneciphylline as the main compound accounting for more than 90% of the alkaloid fraction in all above ground plant parts. Further alkaloids were spartioidine, acetyl-senciphylline and senecionine. Inflorescences showed the highest alkaloid contents with 21.1 and 13.4 mg/g in A. alliariae and A. glabra, respectively. Stems and leaves had 2-3 times lower contents. Therefore, these Adenostyles species must be considered as highly toxic plants.

Pyrrolizidine Alkaloid Production of Jacobaea aquatica under Different Cutting Regimes.

Jacobaea aquatica (Asteraceae) growing in wet grasslands with low management intensity is regarded as a noxious weed with pyrrolizidine alkaloids (PAs), which cause health problems to livestock. The influence of different management practices on the production of PAs and on the proportion of J. aquatica in the fodder was studied. Five cutting regimes were applied during 4 years on permanent plots in lower Austria. The toxicity of the fodder was assessed by recording dry weight and alkaloid content of J. aquatica and total aboveground biomass. Different cutting regimes had significant effects on the PA content of J. aquatica and on its proportion in the fodder. The content of J. aquatica was lowest in fodder of June and October cuts and highest in second cuts in July and August. Total alkaloid contents exceeding 100 mg/kg were found in fodder harvested in July and August. After cutting, the toxic plants regenerated quickly and produced new flowering stalks within 4-5 weeks. Six macrocylic PAs were evaluated, with Z-erucifoline as the most abundant compound. The alkaloid levels were highest in plants cut during summer when flowering plants were present. Consequently, this fodder should not be fed to livestock over a long period of time.

Effects of thyme as a feed additive in broiler chickens on thymol in gut contents, blood plasma, liver and muscle.

BACKGROUND: Aromatic herbs as feed additives in animal production are encountering growing interest, but data on the fate of the aromatic compounds from the plant in the animal body are very scarce. In the present study, thyme (Thymus vulgaris) herb consisting of leaves and flowers without stems was used as an ingredient in the diet for broilers. The herb was fed for 35 days to five groups of broilers (0, 0.1, 0.2, 0.3, and 1% w/w in the diet). Animal performance and the concentrations of the main essential oil component from thyme, thymol, were measured in gut contents, plasma and liver and muscle tissues using solid phase microextraction and gas chromatography/mass spectrometry. RESULTS: There were no differences between the groups in feed intake, daily weight gain, feed conversion and slaughter weight. Thymol was detected in gut contents, plasma and liver and muscle tissues. Increased intestinal thymol concentrations were found in the group with 1% thyme compared with the other groups (P < 0.05). In liver and muscle tissues the thymol levels were close to the limit of quantification. CONCLUSION: The data do not indicate a positive effect of thyme on animal performance. With high dietary levels of thyme herb, thymol concentrations increased in gut contents and plasma but were very low in edible tissues such as liver and flesh. © 2014 Society of Chemical Industry.

Composition of the essential oil of wild grown caraway in meadows of the Vienna region (Austria).

The essential oil compositions of the different plant parts of wild growing caraway (Carum carvi, Apiaceae) were analyzed by GC/MS and GC/FID. The plants came from two meadows in the Vienna region, Austria. The oils from young fruits had a high proportion of limonene (61-83%), and those from inflorescences limonene (39-62%) and germacrene D (23-41%). The carvone contents of fruits, inflorescences and leaves varied greatly. Leaves and stems, low in volatiles, were dominated by germacrene D and germacrene A. Therefore, wild caraway appears as a valuable aromatic herb in meadows.

Chemical characterization by GC-MS and in vitro activity against Candida albicans of volatile fractions prepared from Artemisia dracunculus, Artemisia abrotanum, Artemisia absinthium and Artemisia vulgaris.

BACKGROUND: A large number of essential oils is reported to have significant activity against Candida albicans. But the different chemical composition influences the degree of their activity. The intention of this study was to investigate the chemical composition and the activity against Candida albicans of volatile oils obtained from Artemisia dracunculus, A. abrotanum, A. absinthium and A. vulgaris (Asteraceae). The aim of the study was to identify new chemical compounds that have effect against C. albicans.The essential oils were obtained by hydrodistillation or extraction with dichloromethane (a new procedure we developed trying to obtain better, more separated compounds) from air dried above ground plant material and analyzed by GC-MS. Additionally commercial essential oils from the same species were tested. The Candida albicans inhibition studies were carried out by the paper disc diffusion method. RESULTS: The essential oils shared common components but presented differences in composition and showed variable antifungal activity. Davanone and derivatives thereof, compounds with silphiperfolane skeleton, estragole, davanone oil, ß-thujone, sabinyl acetate, herniarin, cis-chrysanthenyl acetate, 1,8-cineol, and terpineol were the main components of Artemisia volatiles. CONCLUSIONS: Among the volatile fractions tested those from A. abrotanum containing davanone or silphiperfolane derivatives showed the highest antifungal activity. The in vitro tests revealed that the Artemisia oils are promising candidates for further research to develop novel anti-candida drugs.

Composition and variability of the essential oil of Salvia nemorosa (Lamiaceae) from the Vienna area of Austria.

The volatiles present in the aerial parts of Salvia nemorosa L. (Lamiaceae), grave sage, were analysed by GC/MS and GC in plants growing at different sites in the outskirts of Vienna, Austria. The flower oils contained mainly sabinene (37-44%), germacrene D (9-14%), beta-caryophyllene (8-12%) and caryophyllene oxide (2.6-4.4%). Leaf samples had beta-caryophyllene (14-41%), germacrene D (14-38%) and caryophyllene oxide (5-20%) as main compounds, while stem oils were characterized by an high hexadecanoic acid percentage besides germacrene D and beta-caryophyllene.

Composition of the essential oils from Peucedanum cervaria and P. alsaticum growing wild in the urban area of Vienna (Austria).

The composition of the essential oil from the different above ground plant parts of Peucedanum cervaria and P. alsaticum (Apiaceae) collected in the urban area of Vienna has been studied. P. cervaria fruits and inflorescences had 1.5 and 1.8% essential oil, respectively. All the oils of P. cervaria were dominated by monoterpenes, with the main components being beta-pinene (7-58%), alpha-pinene (7-22%), sabinene (up to 22%), and beta-phellandrene with limonene (6-21%). P. alsaticum fruits afforded 0.3-0.4% essential oil. These oils were made up mainly by alpha-pinene (11-40%), sabinene (16-34%) and beta-phellandrene (12-31%). Stems and leaves produced only very small amounts of essential oil (< 0.05%). Besides monoterpenes, these oils contained also E-nerolidol (5-22%), spathulenol (up to 18%), dodecanal (up to 7.5%) and caryophyllene oxide (up to 7%).

Botanical species being used for manufacturing plant food supplements (PFS) and related products in the EU member states and selected third countries.

A great wealth of plants and plant derived preparations are used in the intention to supplement the basic nutrition in order to sustain and promote health. They may be used directly or consumed as manufactured plant food supplements (PFS) in dosed form. The use of these plants may already have a long tradition as fruit, vegetable or (folk) medicinal plants. Due to globalisation, more and more plants originating from all over the world are now offered and marketed in European countries, including species from China, South Africa and the American continent. For reasons of security, EU wide lists of plants accepted or prohibited to be used in food supplements are in elaboration. A crucial point is the correct identification of the plant material. The identity can be assessed by morphological, chemical and DNA specific methods. The active substances usable in PFS are secondary plant products that are often characteristic for certain plant groups (taxa), species or plant parts. They comprise not only polyphenols, essential oils, carotenoids and phytosterols, but also glucosinolates or saponins. The quality of the plant material used for PFS depends on a variety of factors, including the natural phytochemical, intraspecific variation with the occurrence of chemotypes, the ontogenetic variation, the considered plant parts and environmental influences during plant growth. In the production of the raw materials for PFS international standards (good agricultural practice, fair trade) should be applied.

Composition of the essential oils from Anthriscus cerefolium var. trichocarpa and A. caucalis growing wild in the urban area of Vienna (Austria).

The composition of the essential oil from the different above ground plant parts of Anthriscus cerefolium and A. caucalis collected in the urban area of Vienna has been studied. The essential oils of A. cerefolium were dominated by estragole (= methylchavicol) and 1-allyl-2,4-dimethoxybenzene, occurring in various proportions. The oils from young flowering plants had more estragole than 1-allyl-2,4-dimethoxybenzene. Fruits from one location reached 95% estragole in the essential oil. In A. caucalis, the main compound of all oils was cis-chrysanthenyl acetate (up to 74%) followed by cis-chrysanthenol (up to 16%). The fruit oils displayed a higher proportion of cis-chrysanthenyl acetate than the leaf or stem oils.

Essential oil composition of wild growing Apiaceae from Europe and the Mediterranean.

In this overview, the essential oil composition of more than 150 species from about 50 genera of the Apiaceae is reviewed. Essential oil components encountered in Apiaceae proved to be very diverse; they include monoterpenes, sesquiterpenes, rarely diterpenes, phenylpropanoids, phthalides, octanol and octyl esters, trimethylbenzaldehydes and aliphatic aldehydes. In assessing the data one has to keep in mind that the essential oil amount and composition is influenced by many factors, including geographic and genetic variation, physiological aspects and environmental conditions. In some species the occurrence of chemotypes could be demonstrated, but in many cases the data available do not allow the evaluation of infraspecific variability.

Composition at different development stages of the essential oil of four Achillea species grown in Iran.

Four Achillea species, A. millefolium, A. nobilis, A. eriophora and A. biebersteinii, were grown in small field plots in Iran and harvested at four developmental stages: vegetative, at the appearance of the first flower heads, at full flowering, and at late flowering. The composition of the main volatile compounds in dichloromethane extracts and the essential oil obtained by microdistillation was established by GC/MS and GC. 1,8-Cineole (27-41%) was the main compound in the oils from A. millefolium and A. biebersteinii. These two species reached the highest amount of volatile compounds at the full blooming stage. alpha-Thujone was the main compound in A. nobilis oil (25-64%). Fully blooming plants of this species also had a high proportion of artemisia ketone (up to 40%) in the oil. The main oil compounds of A. eriophora were camphor (about 35%) and 1,8-cineol (about 30%). This species produces only a small number of flower heads and the composition of the essential oil did not change during development.