Quillaja Saponaria Saponins Research Articles

INVESTIGATION OF THE POSSIBILITY OF USING THE METHOD OF DIFFERENTIAL SPECTROPHOTOMETRY FOR THE ANALYSIS OF OVERLAPPING ABSORPTION BANDS OF TRITERPENE SAPONINS CHROMOPHORES AND THEIR QUANTITATIVE DETERMINATION IN AQUEOUS SOLUTIONS

A detailed analysis of the absorption spectra of mono- and bidesmoside triterpene glycosides was performed. It is suggested that the maximum in the region of 198–208 nm is attributed to the absorption of the double bond in the cyclohexene ring of the aglycone. The second less seen maximum at a wavelength of 280 nm is observed in the spectrum of saponin Quillaja Saponaria and is almost completely absent in the spectrum of saponin Mukorossi Sapindus. Identification of this maximum is extremely difficult, since its presence can be due to both the aldehyde group in the aglycone and the hydroxyl and carboxyl groups in the carbohydrate molecules.
 Using the method of differential spectrophotometry, a method for decomposing the UV spectrum of saponins into its constituent components is proposed and justified. A spectral analysis was performed, including the assignment of the absorption bands to the functional groups of the studied compounds. The possibility of estimating changes in the spectral properties of glycoside solutions with changes in the acidity of the medium is considered. The possibility of a bathochromic shift of the maximum absorption of the aglyconic part of saponin depending on the microenvironment (the structure of the carbohydrate part) and changes in the pH of the solution is established. It is shown that the bands at 274, 280.5 nm correspond to n-π* transitions of carbonyl and carboxyl groups and are determined only in the regions of high concentrations in Quillaja Saponaria saponin solutions. Based on the results obtained, calibration models for the quantitative determination of saponins in solutions are proposed. The regression analysis of the calibration equations is carried out, the main statistical indicators are calculated.

In vitro efficacy of Quillaja saponaria extracts on the infective life‐stage of ectoparasite Caligus rogercresseyi

AbstractThe effect of two different commercial products of Quillaja saponaria saponin extract on mortality of the free‐living copepodite stage sea‐lice Caligus rogercresseyi were assessed in vitro. Parasites were exposed for 24 hr to different concentrations of Quillaja extracts and then observed until 30 hr in non‐Quillaja media. The EC50 and EC90 were evaluated at 30 hr. High mortalities of copepodites were obtained at higher doses (500 ppm) for both extracts. Higher mortalities were obtained using the product with higher saponin content at 24 hr (23.3% vs. 86.6%) and 30 hr (63.3% vs. 93.3%) compared with the product containing less saponins. In addition, mortalities increased even after removing the extracts suggesting a residual and prolonged effect on the survival of the parasite. Our results indicated that the free‐living stage of the parasite is sensitive to saponins from Q. saponaria extracts and may be used to control or prevent the infestation of C. rogercresseyi in salmonids in Chile.

Quillaja saponaria Saponins with Potential to Enhance the Effectiveness of Disinfection Processes in the Beverage Industry

This study examines the in vitro effect of Quillaja saponaria extracts on Asaia spp. planktonic cells and biofilms, in comparison and combination with two disinfectants: peracetic acid and N-ethyl-N,N-dimethylhexadecylammonium bromide. The growth of six bacterial strains was evaluated spectrophotometrically. Biofilm eradication was determined using the plate count method and luminometry. The planktonic cells were characterized by relatively high resistance to peracetic acid and higher sensitivity to N-ethylo-N,N-dimethylohexadecylioamonium bromide. In almost all the tested strains, growth was inhibited by 0.125% (v/v) peracetic acid and 0.0313% (w/v) quaternary ammonium compound. However, combinations of cell pretreatment using saponin and peracetic acid action were the most efficient against both planktonic and biofilm cells. The minimum inhibitory concentrations for peracetic acid were 4–8 times lower than those for bacterial strains without preliminary saponin action. Eradication of Asaia spp. biofilms reduced the number of living cells by 4–5 logarithmic units. These results demonstrate the synergetic action of saponin extract and disinfectant, and could be useful in the development of industrial strategies against Asaia spp. biofilms.

Purification of an Immunoadjuvant Saponin Fraction from Quillaja brasiliensis Leaves by Reversed-Phase Silica Gel Chromatography.

Saponins include a large variety of molecules that find several applications in pharmacology. The use of Quillaja saponaria saponins as immunological adjuvants in vaccines is of interest due to their capacity to stimulate both humoral and cellular responses. The congener species Q. brasiliensis has saponins with chemical similarities and adjuvant activity comparable to that of Q. saponaria fraction Quil-A®, with additional advantages of showing lower toxicity and reduced hemolytic activity. Here we describe in detail the methods for preparing the aqueous extract from Q. brasiliensis leaves, as well as the purification of the bioactive saponin fraction QB-90 using silica reversed-phase chromatography.

Adjuvant Effect of Quillaja saponaria Saponin (QSS) on Protective Efficacy and IgM Generation in Turbot (Scophthalmus maximus) upon Immersion Vaccination.

The adjuvant effect of Quillaja saponaria saponin (QSS) on protection of turbot fry was investigated with immersion vaccination of formalin-killed Vibrio anguillarum O1 and various concentrations of QSS (5, 25, 45 and 65 mg/L). Fish were challenged at days 7, 14 and 28 post-vaccination. Significantly high relative percent of survival (RPS) ((59.1 ± 13.6)%, (81.7 ± 8.2)%, (77.8 ± 9.6)%) were recorded in the fish that received bacterins immersion with QSS at 45 mg/L, which is comparable to the positive control group vaccinated by intraperitoneal injection (IP). Moreover, a remarkably higher serum antibody titer was also demonstrated after 28 days in the vaccinated fish with QSS (45 mg/L) than those vaccinated fish without QSS (p < 0.05), but lower than the IP immunized fish (p < 0.05). Significant upregulation of IgM gene expression has also been identified in the tissues of skin, gill, spleen and kidney from the immunized fish in comparison to the control fish. Taken together, the present study indicated that QSS was able to dramatically evoke systemic and mucosal immune responses in immunized fish. Therefore, QSS might be a promising adjuvant candidate for fish vaccination via an immersion administering route.

Novel ISCOMs from Quillaja brasiliensis saponins induce mucosal and systemic antibody production, T-cell responses and improved antigen uptake

In the last decades, significant efforts have been dedicated to the search for novel vaccine adjuvants. In this regard, saponins and its formulations as “immunostimulating complexes” (ISCOMs) have shown to be capable of stimulating potent humoral and cellular immune responses, enhanced cytokine production and activation of cytotoxic T cells. The immunological activity of ISCOMs formulated with a saponin fraction extracted from Quillaja brasiliensis (QB-90 fraction) as an alternative to classical ISCOMs based on Quil A® (IQA) is presented here. The ISCOMs prepared with QB-90, named IQB-90, typically consist of 40–50nm, spherical, cage-like particles, built up by QB-90, cholesterol, phospholipids and antigen (ovalbumin, OVA). These nanoparticles were efficiently uptaken in vitro by murine bone marrow-derived dendritic cells. Subcutaneously inoculated IQB-90 induced strong serum antibody responses encompassing specific IgG1 and IgG2a, robust DTH reactions, significant T cell proliferation and increases in Th1 (IFN-γ and IL-2) cytokine responses. Intranasally delivered IQB-90 elicited serum IgG and IgG1, and mucosal IgA responses at distal systemic sites (nasal passages, large intestine and vaginal lumen). These results indicate that IQB-90 is a promising alternative to classic ISCOMs as vaccine adjuvants, capable of enhancing humoral and cellular immunity to levels comparable to those induced by ISCOMs manufactured with Quillaja saponaria saponins.

Evaluation of the anti-inflammatory activities of Quillaja saponaria Mol. saponin extract in mice.

ObjectiveQuillaja saponaria bark contains a high percentage of triterpene saponins and has been used for centuries as antiinflammatory and analgesic agent in Chilean folk medicine.In the Present study the anti-inflammatory activities of the aqueous extract of commercially partially purified saponin from Quillaja saponaria Mol. in in vivo animal models.Methods & materials:Aqueous extract of the plant material was prepared by cold maceration. The anti-inflammatory activity of a commercial Quillaja saponaria Mol. (QS) saponin extract was investigated by carragenan induced mice paw edema model for acute inflammation (Winter, 1962) [16].ResultsThe anti-inflammatory activity was evaluated by carragenan in paw edema model in swiss albino mice (18–20 g). The anti-inflammatory activity was found to be dose dependent in carragenan induced paw edema. QS was found to significantly (p < 0.05) reduce the carragenan induced mice paw edema (38.59%; 20 mg/kg bw) as compared to carragenan control. The percentage inhibition of standard anti-inflammatory drug indomethacin was (55%; 10 mg/kg, bw).ConclusionThe results of the present study demonstrate that the aqueous extract of Quillaja saponaria saponins (QS) possess significant anti-inflammatory activity.

Effects of phytogenic feed additives on growth performance and on ammonia and greenhouse gases emissions in growing-finishing pigs

The present study was conducted to evaluate the effects of two commercial phytogenic feed additives (PFAs) on growth performance and emissions of ammonia, methane, nitrous oxide and carbon dioxide compared to an unsupplemented control. The two tested commercial PFAs contained either essential oils or a mixture of essential oils and dried herbs and spices. In addition, both PFAs delivered the same amount of Quillaja saponaria saponins to the final feed, although dosage of the products differed. The PFAs were included in cereal-based grower and finisher diets via premixes. For negative control diet a placebo premix was used. The diets were fed to a total of 81 castrated male growing-finishing pigs (45–114kg body weight, 27 per treatment) that were kept in nine gas-tight sealed chambers, each with three pens with fully slatted floors (three pigs per pen). The whole experiment lasted 72 days. Pigs were individually weighted and the feed consumption of pigs was recorded per pen at day 0, 24 and 72 of the experiment. From day 24 to day 72 (48 days) emission measurement took place.The inclusion of the PFAs significantly improved average daily feed intake (P=0.010) and average daily gain (P=0.018) of pigs over the whole trial period of 72 days compared to the negative control. Feed conversion ratio was not affected by the treatments (P>0.05). Pigs that were fed with the PFAs had 3.6% higher final body weight (P=0.017) compared to the negative control. The inclusion of the PFAs reduced ammonia emissions per animal per day (P=0.003) as well as per kg body weight gain of pigs (P<0.001) on average by 21% and 26%, respectively, and tended to reduce carbon dioxide emissions per kg body weight gain (P=0.092) on average by 9% compared to the negative control. Methane and nitrous gas emissions were not affected by the inclusion of the PFAs (P>0.05).In summary, it might be speculated that the observed increased intake and consequently higher growth was mediated via flavoring properties of the PFAs, irrespective of differences in composition. The reduction of ammonia emissions most probably was due to the inclusion of the quillaja saponins in the PFAs. It can be concluded that the tested PFAs have a potential as performance enhancers and are useful tools for the reduction of ammonia emissions from pig barns. Further research is warranted to identify the exact modes of action of PFAs.

Cell lysis induced by membrane-damaging detergent saponins from Quillaja saponaria

This paper presents the results of a study to determine the effect of Quillaja saponaria saponins on the lysis of industrial yeast strains. Cell lysis induced by saponin from Q. saponaria combined with the plasmolysing effect of 5% NaCl for Saccharomyces cerevisiae, Kluyveromyces marxianus yeasts biomass was conducted at 50°C for 24–48h. Membrane permeability and integrity of the yeast cells were monitored using fluorescent techniques and concentrations of proteins, free amino nitrogen (FAN) and free amino acids in resulting lysates were analyzed. Protein release was significantly higher in the case of yeast cell lysis promoted with 0.008% Q. saponaria and 5% NaCl in comparison to plasmolysis triggered by NaCl only.

Preparation of human tissue protein extracts enriched with the sphingomyelin synthase 1

Sphingomyelin synthase 1 (SMS 1) catalyzes sphingomyelin biosynthesis in eukaryotic cells. We previously studied the structure of the human SGMS1 gene, which encodes the enzyme and its numerous transcripts. The tissue-specific expression of the transcripts was also described. Analysis of the SMS1 protein expression in human tissues using immunoblotting of tissue extracts prepared in the RIPA (Radio Immuno-Precipitation Assay) buffer revealed a weak signal in renal cortex, testis, lung, and no signal in placenta and lymphatic node. In this work, a new method of preparation of the tissue protein extracts enriched with SMS1 was suggested. The method based on the consecutive extraction with a buffer containing 0.05 and 1 mg/ml of the Quillaja saponaria saponin allowed SMS1 to be detected in all tissues tested. The SMS1 content in the saponin extract of kidney cortex is about 12-fold higher compared to the RIPA extraction procedure.

Association between haptoglobin and IgM levels and the clinical progression of caseous lymphadenitis in sheep

BackgroundSheep caseous lymphadenitis (CLA), caused by Corynebacterium pseudotuberculosis (Cp), is associated with direct economic losses and presents significant zoonotic potential. Despite the importance of the disease, a satisfactory vaccine model has not been developed. Thus, this study aimed to investigate the association between haptoglobin (Hp) and IgM levels and the clinical progression of CLA in primarily infected sheep and in sheep immunized with Cp- secreted antigens adjuvanted with Quillaja saponaria saponins. These animals were kept with CLA-positive sheep to simulate natural exposure that occurs in field conditions. During the experiment, the Hp and IgM levels were monitored for 21 days, and the development of internal CLA lesions was investigated through necropsies on day182 post-immunization.ResultsPrimarily infected sheep in Group 2 (inoculated with 2x105 Cp virulent strain) had higher Hp values between the first and ninth days post inoculation (PI) than sheep in Group 1 (control; P < 0.05). Immunized animals in Group 3 had significantly higher Hp values between the third and seventh days PI, compared with the control group (P < 0.01). Binary logistic regression (BLR) analysis of primarily infected sheep indicated an association between Hp concentration and CLA clinical progression: animals with high Hp values had 99.9% less risk of having CLA abscesses than animals with low Hp levels (Odds ratio = 0.001, P < 0.05). Both experimental groups had significantly higher IgM titers than the control group around the ninth and eleventh days PI (P < 0.05). The BLR analysis for immunized sheep indicated an association between IgM levels and clinical progression: sheep with high IgM titers had 100.0% less risk of having CLA abscesses than animals with low IgM levels (Odds ratio = 0.000, P < 0.05).ConclusionsResistance to C. pseudotuberculosis infection is supported by the early acute phase response, in which up-regulation of Hp and IgM were predictive of a lower risk of CLA lesion development. Because the immunogen used in this study induced a high production of both Hp and IgM, Q. saponaria saponin should be considered a promising candidate in vaccine formulations against sheep CLA.

Saponins do not affect the ecdysteroid receptor complex but cause membrane permeation in insect culture cell lines

This project studied the effects of four saponins with a triterpenoid (Quillajasaponaria saponin and aescin) or steroid structure (digitonin and diosgenin which is the deglycosylated form of dioscin) on insect cells, namely Schneider S2 cells of Drosophila melanogaster (Diptera). A series of different experiments were performed to investigate potential mechanisms of action by saponins with regard to ecdysteroid receptor (EcR) responsiveness, cell viability, cell membrane permeation, and induction of apoptosis with DNA fragmentation and caspase-3 like activity.Major results were that (1) exposure of S2 cells containing an EcR-based reporter construct to a concentration series of each saponin scored no EcR activation, while (2) a loss of ecdysteroid signaling was observed with median inhibitory concentrations (IC50’s) of 3–50μM, and in parallel (3) a concentration-dependent change in loss of cell numbers in an cell viability assay with median effective concentrations (EC50’s) of 8–699μM. In continuation, it was of interest that (4) a trypan blue assay with Q. saponaria saponin confirmed the cell membrane permeation effect leading to cell toxicity with a median lethal concentration (LC50) value of 44μM, and interestingly this effect was very rapid. Another three interesting observations were that (5) exposure to 20E at 500nM as used in the EcR-based report assay induced caspase-3 like activities which may help to explain the discrepancies between loss of EcR-responsiveness and cell viability, (6) low concentrations of saponins induced DNA fragmentation and caspase-3 like activities, confirming their potential to induce apoptosis, and (7) the saponin effects were counteracted with addition of cholesterol to the culture medium.In general the data obtained provide evidence that the anti-ecdysteroid action by saponins is not based on a true antagonistic interaction with EcR signaling, but can be explained by a cytotoxic action due to permeation of the insect cell membrane.

Impact of Quillaja saponaria saponins on grapevine ecosystem organisms

The control of grapevine pathogens is a rising concern in Vitis vinifera culture. The current international trend is toward banning chemicals that are highly toxic to the environment and human workers, and adopting tighter regulations. We evaluated the impact of saponins on three kinds of organisms found in grapevine culture. The ectoparasitic nematode Xiphinema index, the parasitic fungus Botrytis cinerea and various yeast strains representative of the must fermentation population were incubated on synthetic media supplemented with variable concentrations of Quillaja saponaria saponins. Saponins induced reduction in the growth of B. cinerea and showed nematicide effects on X. index. The control of X. index and Botrytis cinerea is discussed in the context of the potential use of these chemicals as environmentally-friendly grapevine treatments. With Saccharomyces cerevisiae and other yeasts, saponins showed higher toxicity against S. cerevisiae strains isolated from wine or palm wine whereas laboratory strains or strains isolated from oak exhibited better resistance. This indicates that Q. saponaria saponins effects against yeast microflora should be assessed in the field before they can be considered an environmentally-safe new molecule against B. cinerea and X. index.

Antinociceptive activity of Quillaja saponaria Mol. saponin extract, quillaic acid and derivatives in mice

Quillaja saponaria bark contains a high percentage of triterpene saponins and has been used for centuries as a cleansing and analgesic agent in Chilean folk medicine. The topical and systemic analgesic effects of a commercial partially purified saponin extract, 3β,16α-dihydroxy-23-oxoolean-12-en-28-oic acid (quillaic acid), methyl 3β,16α-dihydroxy-23-oxoolean-12-en-28-oate and methyl 4-nor-3,16-dioxoolean-12-en-28-oate. The samples were assessed in mice using the topical tail-flick and i.p. hot-plate tests, respectively. All the samples showed activity in both analgesic tests in a dose-dependent manner. The most active against tail flick test was commercial partially purified saponin extract (EC50 27.9 mg%, w/v) and more than the ibuprofen sodium. On hot-plate test, methyl 4-nor-3, 16-dioxoolean-12-en-28-oate was the most active (ED50 12.2 mg/kg) and more than the ibuprofen sodium. The results of the present study demonstrated that Quillaja saponaria saponins, quillaic acid, its methyl ester, and one of the oxidized derivatives of the latter, elicit dose-dependent antinociceptive effects in two murine thermal models.

Post-Plant Nematicides for the Control of Root Lesion Nematode in Red Raspberry

To identify a post-plant nematicide to control root lesion nematode [RLN (Pratylenchus penetrans)] in red raspberry (Rubus idaeus), a number of nematicides was tested in soil-only and plant-based experiments. In soil-only experiments, soil naturally infested with RLN was drenched with the nematicides and nematode survival was assessed 7 and 14 days after treatment. Fosthiazate and oxamyl reduced RLN recovery 92% and 52% across trials and sampling times, respectively, compared with the nontreated control. Other nematicides that resulted in moderate, and sometimes inconsistent, control of RLN were soapbark (Quillaja saponaria) saponins, 1,3-dichloropropene, and methomyl. In plant-based experiments, ‘Meeker’ red raspberry was established in pots with RLN-infested soil mixed with greenhouse soil and the nematicides were applied as soil drenches or as a foliar application. Nematode recovery and cane and root weights were quantified as measurements of nematicide toxicity and phytotoxicity, respectively. Similar to soil-only experiments, fosthiazate and oxamyl were the most effective nematicides tested in reducing RLN population densities in established red raspberry. Fosthiazate and oxamyl significantly reduced RLN per gram dry root population densities by 97% and 87%, respectively, compared with the infested, nontreated control. None of the other nematicides reduced RLN population densities compared with the infested, nontreated controls. There was no phytotoxicity to red raspberry associated with any of the nematicides.

Quillaja saponaria saponin is causing an anti-ecdysteroid action in insect cells that may be explained by cytotoxicity and permeation

Event Abstract Back to Event Quillaja saponaria saponin is causing an anti-ecdysteroid action in insect cells that may be explained by cytotoxicity and permeation Guy Smagghe1*, Ellen De Geyter1, 2, Danny Geelen1, Thomas Soin1 and Luc Swevers3 1 Ghent University, Department of Crop Protection, Faculty of Bioscience Engineering, Belgium 2 Ghent University, Department of Plant Production, Faculty of Bioscience Engineering, Belgium 3 National Centre of Scientific Research ‘Demokritos’, Institute of Biology, Greece In this project we studied the cytotoxic effects of the triterpenoid (five-ringed) saponin of the oleanane-glucuronide-type from the soapbark tree Quillaja saponaria. In a first series of experiments, we transfected S2 observed a concentration dependence for the ecdysteroid responsiveness of the cells when these were transfected with an EcR-reporter construct. There was no activation of the EcR-signaling, but we demonstrated a typical loss of ecdysteroid signaling at low concentrations with a respective IC50 of 1.7 µM and 0.68 µM. A concentration-dependent change in cell survival was observed when insect cells of Drosophila melanogaster (S2, embryo) and Bombyx more (Bm5, ovary) were incubated. A loss of 50% of cell survival (LC50) was observed at 5.1 µM and 1.7 µM, respectively, in an MTT bioassay. The cell permeation was also confirmed in a trypan blue assay. To explain the latter anti-ecdysteroid action we investigated be explained by the cytotoxic and permeation action. In addition, it was of interest that saponin effects were counteracted with addition of cholesterol to the cell culture medium. Finally, caspase 3-like measurements showed that Quillaja saponin could not induce nuclear events as apoptosis in treated cells. Our results suggest that the loss of ecdysteroid responsiveness after the addition of Quillaja saponin could thus be result of a cytotoxic permeation action and not by interacting with the EcR. Acknowledgements This work was supported by the Special Research Fund of Ghent University, and the Fund for Scientific Research (FWO-Vlaanderen, Brussels) Keywords: comparative endocrinology Conference: 25th Conference of the European Comparative Endocrinologists, Pécs, Hungary, 31 Aug - 4 Sep, 2010. Presentation Type: Conference Presentation Topic: Comparative endocrinology Citation: Smagghe G, Geyter E, Geelen D, Soin T and Swevers L (2010). Quillaja saponaria saponin is causing an anti-ecdysteroid action in insect cells that may be explained by cytotoxicity and permeation. Front. Endocrinol. Conference Abstract: 25th Conference of the European Comparative Endocrinologists. doi: 10.3389/conf.fendo.2010.01.00063 Copyright: The abstracts in this collection have not been subject to any Frontiers peer review or checks, and are not endorsed by Frontiers. They are made available through the Frontiers publishing platform as a service to conference organizers and presenters. The copyright in the individual abstracts is owned by the author of each abstract or his/her employer unless otherwise stated. Each abstract, as well as the collection of abstracts, are published under a Creative Commons CC-BY 4.0 (attribution) licence (https://creativecommons.org/licenses/by/4.0/) and may thus be reproduced, translated, adapted and be the subject of derivative works provided the authors and Frontiers are attributed. For Frontiers’ terms and conditions please see https://www.frontiersin.org/legal/terms-and-conditions. Received: 27 Aug 2010; Published Online: 29 Aug 2010. * Correspondence: Dr. Guy Smagghe, Ghent University, Department of Crop Protection, Faculty of Bioscience Engineering, Ghent, Belgium, guy.smagghe@ugent.be Login Required This action requires you to be registered with Frontiers and logged in. To register or login click here. Abstract Info Abstract The Authors in Frontiers Guy Smagghe Ellen De Geyter Danny Geelen Thomas Soin Luc Swevers Google Guy Smagghe Ellen De Geyter Danny Geelen Thomas Soin Luc Swevers Google Scholar Guy Smagghe Ellen De Geyter Danny Geelen Thomas Soin Luc Swevers PubMed Guy Smagghe Ellen De Geyter Danny Geelen Thomas Soin Luc Swevers Related Article in Frontiers Google Scholar PubMed Abstract Close Back to top Javascript is disabled. Please enable Javascript in your browser settings in order to see all the content on this page.

Immunostimulatory characteristics of a novel adjuvant on the basis of cucumarioside A2-2 and monogalactosyldiacylgycerol

A novel antigen carrier has been formulated on the basis of a cucumarioside-A2-2 triterpene glycoside (CD) complex with cholesterol and monogalactosyldiacylglycerol from Ahnfeltia tobuchiensis (MGDG(At)) and Ulva fenestrate (MGDG(Uf)). Morphological and immunostimulative characteristics of the carrier were studied. Electron microscopy experiments demonstrated the formation of homogeneous tubular structures in a mixture of CD, cholesterol, and MGDG in molar ratio of 1:2:3. In animals immunized by the carrier bearing pore forming protein monomer of pseudotuberculosis agent CD and MGDG synergically affected synthesis of specific antibodies, interleukin-2, and gamma-interferon and delayed hypersensitivity reaction when compared to Freund's complete adjuvant or to immunostimulatory complexes between Quillaja saponaria saponins and phosphatidylcholine from egg yolk. The immunostimulatory effect depends upon the composition of polyunsaturated fatty acids of MGDG. The new tubular adjuvant carrier is a competitive adjuvant, as it includes CD obtained from far-eastern sea cucumber commercial species Cucumaria japonica, and MGDG from seaweed.

Adjuvant activities of saponins from the root of Polygala senega L.This article is one of a selection of papers published in this special issue (part 2 of 2) on the Safety and Efficacy of Natural Health Products.

Eight pure triterpenoid saponin compounds isolated from the root of Polygala senega L., a plant indigenous to the Canadian prairies, were evaluated for their immunological activity in mouse models. The specific antibody responses of the IgG2a subclass increased significantly when isolated P. senega saponins were used as adjuvants in the immunization of mice with OVA antigen. In addition, increased IL-2 levels were observed in spleen cell cultures from P. senega saponin-immunized mice after in vitro secondary antigen stimulation. The saponins were tested for their toxicity in mice by using a haemolytic activity assay and found to be less toxic than Quillaja saponaria saponins that have long been used as adjuvants in vaccine formulations. This study has shown the potential of P. senega saponins to be considered as a natural source of vaccine adjuvants with biological activity equivalent to the current commercially available saponin adjuvants.

Differences in the in vitro susceptibility of mono-eukaryotic cultures of Histomonas meleagridis, Tetratrichomonas gallinarum and Blastocystis sp. to natural organic compounds

Currently, all pharmaceuticals for the treatment or prophylaxis of blackhead disease (histomonosis) caused by the flagellate Histomonas meleagridis are banned from the market. Consequently, great interest exists on the finding of alternative drugs for the abatement of histomonosis. In this study, carvacrol, Cassia oil, an essential oil (EO) mixture containing thyme and rosemary EO and a Quillaja saponaria saponin were examined using in vitro assays for antiprotozoal and antibacterial activity testing established against cloned xenic cultures of different isolates of Histomonas meleagridis, Tetratrichomonas gallinarum and Blastocystis sp. Whereas similar minimal lethal concentrations (MLCs) of five Histomonas isolates were obtained for both carvacrol and the EO mixture as well as for the saponin, significantly different MLCs were observed for them with Cassia oil, ranging from 0.25 up to 0.50 microl/ml. Testing the Blastocystis isolates, different MLCs were obtained for all substances, whereas the Tetratrichomonas gallinarum isolates showed identical susceptibilities. The effects are independent of the bacteria, underlining the need of well-defined protozoan cultures for these investigations.

Adjuvant activity of Quillaja brasiliensis saponins on the immune responses to bovine herpesvirus type 1 in mice

The chemical characterization of aqueous extracts (AE) of barks, leaves and branches and the saponin fraction denominated QB-90 obtained from Quillaja brasiliensis, a native species from Southern Brazil, show remarkable similarities to Quillaja saponaria saponins which are known as adjuvants in vaccine formulations . In vivo toxicity assays of AE and QB-90 showed not to be lethal for mice in doses ranging from 50 to 1600 μg and 50–400 μg, respectively. Experimental vaccines prepared with bovine herpesvirus type 1 (BHV-1) antigen and either AE (barks 100 μg, leaves 400 μg, branches 400 μg) or QB-90 (100 μg) were able to enhance the immune responses of mice in a comparable manner to saponins from Q. saponaria (QuilA, 100 μg) . BHV-1 specific IgG, IgG1 and IgG2a antibody levels in serum were also significantly enhanced by AE, QB-90 and QuilA compared to control group ( p < 0.05). These results showed that AE and QB-90 from Q. brasiliensis are potential candidates as adjuvants in vaccines.