Petr Marsik

Production of saponins from Panax ginseng suspension and adventitious root cultures

Biomass growth and ginsenoside production in cell suspension and adventitious roots of Panax ginseng C.A. Meyer cultures cultivated both in Erlenmayer flasks and a 3 dm3 bioreactor were studied. The maximum content of ginsenosides was found in the suspension culture cultivated in the bioreactor (4.34 % dry mass), however the saponin content was limited to two major ginsenosides, Rb1 and Rg1. The production of ginsenosides in adventitious roots was lower (1.45 or 1.72 % dry mass), nevertheless, the full range of ginsenosides was detected.

Growth of bifidobacteria and clostridia on human and cow milk saccharides.

For healthy infants, which were born normally and fully breastfed, the dominant component of the intestinal microflora are bifidobacteria. However, infants born by caesarean section possess clostridia as a dominant intestinal bacterial group. The aim of the present study was to determine whether bifidobacteria and clostridia are able to grow on human milk oligosaccharides (HMOs) and other carbon sources - lactose, cow milk (CM) and human milk (HM). Both bifidobacteria and clostridia grew on lactose and in CM. Bifidobacteria grew in HM and on HMOs. In contrast, 3 out of 5 strains of clostridia were not able to grow in HM. No clostridial strain was able to utilise HMOs. While both bifidobacterial strains were resistant to lysozyme, 4 out of 5 strains of clostridia were lysozyme-susceptible. It seems that HMOs together with lysozyme may act as prebiotic-bifidogenic compounds inhibiting intestinal clostridia.

Opposing effects of oxidative challenge and carotenoids on antioxidant status and condition-dependent sexual signalling.

Several recent hypotheses consider oxidative stress to be a primary constraint ensuring honesty of condition-dependent carotenoid-based signalling. The key testable difference between these hypotheses is the assumed importance of carotenoids for redox homeostasis, with carotenoids being either antioxidant, pro-oxidant or unimportant. We tested the role of carotenoids in redox balance and sexual signalling by exposing adult male zebra finches (Taeniopygia guttata) to oxidative challenge (diquat dibromide) and manipulating carotenoid intake. As the current controversy over the importance of carotenoids as antioxidants could stem from the hydrophilic basis of commonly-used antioxidant assays, we used the novel measure of in vivo lipophilic antioxidant capacity. Oxidative challenge reduced beak pigmentation but elicited an increase in antioxidant capacity suggesting resource reallocation from signalling to redox homeostasis. Carotenoids counteracted the effect of oxidative challenge on lipophilic (but not hydrophilic) antioxidant capacity, thereby supporting carotenoid antioxidant function in vivo. This is inconsistent with hypotheses proposing that signalling honesty is maintained through either ROS-induced carotenoid degradation or the pro-oxidant effect of high levels of carotenoid-cleavage products acting as a physiological handicap. Our data further suggest that assessment of lipophilic antioxidant capacity is necessary to fully understand the role of redox processes in ecology and evolution.

Hydroxylated Anthraquinones Produced by Geosmithia species

Geosmithia fungi are little known symbionts of bark beetles. Secondary metabolites of lilac colored species G. lavendula and other nine Geosmithia species were investigated in order to elucidate their possible role in the interactions of the fungi with environment. Hydroxylated anthraquinones (yellow, orange, and red pigments), were found to be the most abundant compounds produced into the medium during the submerged cultivation. Three main compounds were identified as 1,3,6,8-tetrahydroxyanthraquinone (1), rhodolamprometrin (1-acetyl-2,4,5,7-tetrahydroxyanthraquinone; 2), and 1-acetyl-2,4,5,7,8-pentahydroxyanthraquinone (3). Compounds 2 and 3 (representing the majority of produced metabolites) inhibited the growth of G+-bacteria Staphylococcus aureus and Bacillus subtilis with minimum inhibitory concentration of 64–512 μg/mL. Anti-inflammatory activity detected as inhibition of cyclooxygenase-2 was found only for compound 3 at 1 and 10 μg/mL. Compound 2 interfered with the morphology, compound 3 with cell-cycle dynamics of adherent mammalian cell lines.

In vitro anti-microbial activity of extracts from the callus cultures of some Nigella species

Crude methanol extracts from callus cultures of Nigella arvensis, N. damascena, N. hispanica, N. integrifolia, and N. sativa were investigated for their anti-microbial activity. Growth inhibition was determined in Gram-positive and Gram-negative bacterial strains as well as in yeast by using a broth-microdilution method. The results showed that the extracts of all calli tested exhibited significant anti-microbial activity, especially against Bacillus cereus, Staphylococcus aureus and Staphylococcus epidermidis. Compared with other Nigella species, a callus culture of N. hispanica was the most effective against the microorganisms used in this study.

The health signalling of ornamental traits in the Grey Partridge (Perdix perdix)

AbstractBirds express various secondary ornaments that can indicate individual condition and health. Amongst these, red-coloured carotenoid-based ornaments are supposed to be particularly valuable predictors of individual quality, due to their sensitivity to oxidative stress. Nevertheless, melanin-pigmented traits might also signal health and immune functions. Both types of ornaments may be either skin-based or feather-based, each differing in their dynamics. In the present study, we compared the health- and stress-indicating capacity of melanin-based feather ornamentation and putatively carotenoid-based skin ornamentation in a single species—the Grey Partridge (Perdix perdix), a vulnerable avian species of the European agricultural landscape. In captive males, we firstly verified the carotenoid content of the red-coloured skin tissue behind the eye by chromatography (HPLC). Secondly, we assessed the individual health status of all males by examining differential leukocyte count, the frequency of immature erythrocytes, malaria prevalence and proinflammatory immune responsiveness to phytohaemagglutinin (PHA). Both the size of the melanin-based ornament and red chroma of the carotenoid-based ornament were related to the heterophil:lymphocyte (H/L) ratio. Hence, in the Grey Partridge, both redness of the skin ornament and area of the feather ornament may serve as honest indicators of individual health and long-term stress. However, the two ornamental components were unrelated to each other, and the directions of their association to the H/L ratio were opposite. We therefore propose that, in this species, larger melanin-based feather ornamentation size is linked to higher levels of stress (possibly caused by more intensive social interactions with other males), while the level of expression of the carotenoid-based skin ornamentation more reliably signals actual individual health status. Our results are potentially valuable from the perspective of Grey Partridge conservation efforts, as they indicate a simple method for assessing individual quality in this species.ZusammenfassungOrnamentmerkmale als Signale für den Gesundheitszustand beim Rebhuhn (Perdix perdix)n Vögel schmücken diverse sekundäre Merkmale, die die Kondition und den Gesundheitszustand eines Individuums anzeigen können. Aufgrund ihrer Empfindlichkeit gegenüber oxidativem Stress gelten unter diesen rote, auf Karotinen basierende Schmuckmerkmale als besonders wichtig. Dennoch können auch auf Melaninpigment basierte Merkmale Gesundheitszustand und Immunfunktionen signalisieren. Beide Ornamenttypen treten sowohl in der Haut als auch in Federn auf, wo sie sich hinsichtlich der Dynamik unterscheiden. In der vorliegenden Arbeit verglichen wir die Kapazität von melaninbasierten Gefiedermerkmalen und die vermutlich karotinbasierte Hautfärbung hinsichtlich ihrer Gesundheitszustands- und Stressindikation beim Rebhuhn (Perdix perdix), einer gefährdeten Vogelart der europäischen Agrarlandschaft. Von Rebhuhnmännchen aus Gefangenschaft bestimmten wir zunächst mittels Chromatographie (HPLC) den Karotingehalt der rot gefärbten Haut hinter dem Auge. Daraufhin ermittelten wir den individuellen Gesundheitszustand aller Männchen indem wir differenzielles Blutbild, den Anteil unreifer Erythrozyten, Malaria-Prävalenz und die entzündliche Immunantwort auf Phytohämagglutinin (PHA) untersuchten. Sowohl die melaninbasierte Ornamentik, als auch die Rotfärbung der auf Karotin basierten Merkmale waren mit dem Verhältnis Heterophile zu Lymphozyten (H/L) korreliert. Damit können beim Rebhuhn beide Schmuckkomponenten, die Rotintensität der Haut, und Gefiederpartien als ehrliche Indikatoren der individuellen Gesundheit und von Langzeitstress dienen. Allerdings ließ sich kein Korrelation zwischen den beiden Komponenten feststellen, und die Zusammenhänge beider mit dem H/L Verhältnis waren gegenteilig. Wir vermuten daher, dass beim Rebhuhn größere melaninbasierte Gefiederornamente mit höheren Stresswerten in Verbindung stehen (möglicherweise verursacht durch intensivere soziale Interaktionen mit anderen Männchen), wohingegen die Stärke der karotinbasierten Hautornamentik zuverlässiger den individuellen Gesundheitsstatus signalisieren. Unsere Ergebnisse sind potentiell aus der Sicht von Rebhuhnschutzmaßnahmen von Nutzen, da sie eine einfache Methode zur Bestimmung individueller Qualität bei dieser Art aufzeigen.

Inter-species differences in the growth of bifidobacteria cultured on human milk oligosaccharides

Human milk (HM) contains as the third most abundant component around 200 different structures of human milk oligosaccharides (HMOs). HMOs are the first and irreplaceable prebiotics for infants, supporting bifidobacteria as the most important bacterial group in an infant intestine. The aim of our study was to test the growth of bifidobacteria in HM and on HMOs. Bifidobacteria were isolated from two groups of infants. The first one (eight strains) were isolated from infants who had bifidobacteria in their feces but, after a short period of time (4 to 24xa0days), bifidobacteria were no longer detected in their feces (disappeared bifidobacteria [DB]). The second group of bifidobacteria (eight strains) originated from infants with continual presence of bifidobacteria in their feces (persistent bifidobacteria [PB]). There were significant differences (pu2009<u20090.05) between DB and PB groups in the ability of the strains to grow in HM. PB grew in HM, reaching counts higher than 7 log CFU/ml. In contrast, counts of DB decreased from 5 to 4.3 log CFU/ml after cultivation in HM. The final pH after cultivation of bifidobacteria on HMOs was 6.2 and 4.9 in DP and PB groups, respectively. In general, Bifidobacterium bifidum and B. breve species were able to utilize HMOs, while B. adolescentis and B. longum subsp. longum species did not. The ability to grow in HM and to utilize HMOs seem to be important properties of bifidobacteria which are able to colonize infant intestinal tract.

Optimalization of the peroxidase production by tissue cultures of horseradish in vitro

Tissue cultures of Armoracia rusticana L., both transformed with Agrobacterium rhizogenes and nontransformed, were screened for peroxidase activity. Most of the derived and tested strains exhibited 20 times higher activity [from 99 to 723 U g−1(d.m.)] than the root of the intact plant [(30 U g−1 (d.m.)]. The highest peroxidase activity was found in tumour culture growing on the medium without growth regulators. The influence of the addition of sugars and heavy metal ions in the medium on peroxidase production was tested. Increase in peroxidase activity was observed after cultivation of horseradish culture with cadmium, cobalt, nickel or lead ions.

Cultivation of root cultures of Panax ginseng in different bioreactors and in temporary immersion — Comparison of growth and saponin production

Different systems of large-scale cultivation of multiple adventitious roots of Panax ginseng C. A. Meyer were compared to cultivation in Erlenmeyer flasks. Adventitious roots were isolated from plantlets regenerated from somatic embryos and cultivated separately in liquid media. Multiplication of adventitious roots was performed in liquid Schenk and Hildebrandt (1972) medium containing 3% sucrose, and 24.6 µmol indole-3-butyric acid. The highest saponin content of 28.51 mg g−1 of dry weight was found in adventitious roots cultivated in the RITA® temporary immersion system (TIS). The best production of biomass was achieved in RITA® vessels and standard Erlenmeyer flasks placed on rotary shaker, followed by the Applikon 3-litre bioreactor and a simple airlift reactor. Saponin production in Erlenmeyer flasks was 10.07 mg g−1 of the dry weight while the production in the Applikon 3-litre bioreactor was only 3.60 mg g−1. Other bioreactor systems tested showed neither significant saponin production nor high biomass production.