Electroantennographic and coupled gas chromatographic-electroantennographic responses of the mediterranean fruit fly,Ceratitis capitata,to male-produced volatiles and mango odor

We have identified five compounds from the headspace of calling male Mediterranean fruit flies (medfly),Ceratitis capitata (Wiedemann), and three compounds from the headspace of ripe mango (Mangifera indica L). using coupled gas chromatographic-electroantennographic (GC-EAG) recordings, coupled gas chromatographic-mass spectrometric (GC-MS) analysis, and electroantennographic (EAG) assays of standards. The male-produced volatiles eliciting responses from female antennae were ethyl-(E)-3-octenoate, geranyl acetate, (E,E)--farnesene, linalool, and indole. An EAG dose-response test of linalool enantiomers and indole with female medfly antennae showed relatively strong EAG activities, but no significant difference between (R)-(-)-linalool and (S)-(+)-linalool. The three mango volatiles were identified as (1S)-(-)--pinene, ethyl octanoate, and-caryophyllene. In addition, a strong antennal response was recorded from a contaminant,-copaene, present in a commercial sample of-caryophyllene. The EAG response amplitudes from both male and female antennae to the above three mango volatiles were significantly greater than to a hexanol control. For both male and female medfly antennae, the greatest EAG responses were elicited by-caryophyllene followed by ethyl octanoate. The mean EAG responses of female antennae to-caryophyllene and (1S)-(-)--pinene were significantly greater than those of male antennae.     

Identification of Volatile Potato Sesquiterpenoids and Their Olfactory Detection by the Two-spotted Stinkbug Perillus bioculatus

The predaceous stinkbug Perillus bioculatus is attracted towards volatiles emitted by damaged potato plants. Whereas mechanically damaged plants lost attractiveness 1 h after damage was inflicted, attraction was long-lasting when the plants were damaged by Colorado potato beetles Leptinotarsa decemlineata, a prey of P. bioculatus. A range of sesquiterpenoids was previously shown to be induced upon beetle damage. In order to evaluate the potential role of sesquiterpenoids in the attraction response, volatiles from damaged potato plants were collected and analyzed with GC-MS and GC-EAG. The antennae of P. bioculatus responded to -caryophyllene, -humulene, (E)--farnesene, (–)-germacrene D, and germacrene D-4-ol. Two sesquiterpenes that coeluted, -zingiberene and bicyclogermacrene, together also elicited olfactory responses of P. bioculatus, whereas the individual compounds did not. The response of P. bioculatus to a variety of sesquiterpenes at low dosages suggests a role for these compounds in prey detection of this stinkbug.     

A Novel Approach for Isolation of Volatile Chemicals Released by Individual Leaves of a Plant in situ

A glass chamber designed specifically for collecting volatile chemicals from individual leaves of a plant in situ is described. The effectiveness of the chamber was demonstrated by collecting volatile chemicals from single leaves of two plant species, potato (Solanum tuberosum) and broad bean (Vicia faba), before and after mechanical damage. The glass chamber, in conjunction with thermal desorption, enables reduction of the entrainment time and thereby allows the monitoring of compounds released by leaf damage in successive 5-min periods. An intact broad bean leaf, in the middle of the day, produces small amounts of the green leaf volatiles (E)-2-hexenal and (Z)-3-hexen-1-ol. However, during the first 5 min after mechanical damage, large amounts of (Z)-3-hexenal, (E)-2-hexenal, and (Z)-3-hexen-1-ol are produced. The decline in production of (Z)-3-hexenal and (E)-2-hexenal is fast, and after 10 min, these compounds reach very low levels. (Z)-3-Hexen-1-ol shows an increase for the first 10 min and then a gradual decline. An intact potato leaf, in the middle of the day, produces very small amounts of the sesquiterpene hydrocarbons -caryophyllene and germacrene-D. After being damaged, the profile of released volatiles is different from that of broad bean. In potato, damage is associated with release of large amounts of green leaf volatiles and sesquiterpene hydrocarbons. Compounds such as (Z)-3-hexenal, (E)-2-hexenal, and (Z)-3-hexen-1-ol are released in high amounts during the first 5 min after damage, but after 10 min, these drop to very low levels. High release associated with damage is also observed for -caryophyllene, (E)--farnesene, germacrene-D, and -bisabolene. The highest level is reached 5 min after damage and 15 min later, these compounds drop to low levels. The significance of compounds released after plant damage is discussed.     

Olfactory Reactions of the Twelve-Spotted Lady Beetle, Coleomegilla maculata and the Green Lacewing, Chrysoperla carnea to Semiochemicals Released from Their Prey and Host Plant: Electroantennogram and Behavioral Responses

Electroantennograms (EAGs) were recorded from two predatory insect species, the twelve-spotted lady beetle, Coleomegilla maculata and the green lacewing, Chrysoperla carnea in response to semiochemicals emitted from one of their prey species, the pea aphid Acyrthosiphon pisum and their host plant. EAG responses were also recorded from C. maculata in response to extracts from individuals of the opposite sex and to extracts from an herbaceous plant, catnip Nepeta cataria. Extracts of catnip and two sex pheromone components of aphids, (4aS,7S,7aR)-nepetalactone and (1R,4aS,7S,7aR)-nepetalactol, elicited significant EAG responses from the antennae of both predatory species. Of 10 corn volatile compounds tested, C. carnea adults responded most strongly to 2-phenylethanol and (E)--farnesene. A significant difference in EAG response to extracts of corn leaf collections was observed between male and female C. carnea. In C. maculata, significant EAG responses were elicited by most of the tested corn volatile compounds, except -pinene and (E)-2-hexenal. The highest EAG responses were observed in response to (E)--farnesene, -terpineol, 2-phenylethanol, and -caryophyllene. Sexual differences in EAG responses of C. maculata were only found in response to 1-octen-3-ol. Male antennae of C. maculata produced significant EAG responses to extracts from conspecific females, but not to males, which indicates that some chemicals from females could be involved in sexual communication. A significant EAG response also was recorded in response to the extracts of fluids produced during reflex bleeding. Male and female antennae of both species exhibited similar dose–response curves to most of the selected compounds, although female C. maculata antennae exhibited higher thresholds in response to several compounds including -terpineol, (Z)-3-hexenol, and (4aS,7S,7aR)-nepetalactone. Field tests showed that 2-phenylethanol was highly attractive to both sexes of the two investigated species. Only C. maculata was attracted to traps baited with -terpineol.     

Electrophysiological Responses of Female and Male Hypsipyla grandella (Zeller) to Swietenia macrophylla Essential Oils

The mahogany shoot borer, Hypsipyla grandella, is one of the most economically important Neotropical forest pest insects because it attacks valuable timber of the Meliaceae. Despite detailed information on the life cycle of H. grandella, factors influencing attack by the shoot borer are not well known. Electroantennogram studies of borer antennae indicate that both female and male moths elicited significant antennal responses to essential oil of the terminal shoots, mature and senescent leaves, in three concentrations. -Himachalene, germacrene D, germacrene A, cadina-1,4-diene, hexadecanoic acid, and ethyl hexadecanoate appeared in all essential oils from Swietenia macrophylla. Germacrene-D and -himachalene were also present in the active essential oil of Cedrela odorata and Toona ciliata. These findings suggest that these compounds play a role in attracting H. grandella to oviposit on the leaves of these plants. The GC-EAD of essential oil from S. macrophylla mature leaves to female H. grandella antennae led to the identification of -caryophyllene as the main constituent responsible for the antennae response.     

Enantiomeric Composition of Monoterpene Hydrocarbons in Some Conifers and Receptor Neuron Discrimination of α-Pinene and Limonene Enantiomers in the Pine Weevil, Hylobius abietis

The enantiomeric composition of seven monoterpene hydrocarbons in headspace volatiles of spruce sawdust and seedlings (Picea abies), pine seedlings (Pinus sylvestris), and branches of juniper (Juniperus communis) was determined by gas chromatographic separation on a -Cyclodextrin column. For the six monoterpenes, -pinene, camphene, -pinene, sabinene, limonene, and -phellandrene, both enantiomers were present, whereas for 3-carene only the (+)-configuration was found. The amount of each enantiomer varied considerably both in relation to total amount of all of them, and for the six pairs also in relation to the opposite enantiomer. One olfactory receptor neuron in the pine weevil (Hylobius abietis) showed a strong response to -pinene when stimulated with all four headspace materials via a GC equipped with a DB-WAX column. The same neuron was subsequently tested with repeated stimulations via the GC effluent containing the (+)- or (–)-enantiomer. A marked better response to (+)- than to (–)--pinene was elicited. Another olfactory receptor neuron that responded strongly to limonene when stimulated with the spruce volatiles was tested for enantiomers of limonene. This neuron responded more strongly to (–)- than to (+)-limonene, when stimulated alternately with each of the limonene enantiomers. Discrimination between enantiomers by plant olfactory receptor neurons suggests that the enantiomeric ratios of volatile compounds may be important in host location by the pine weevil.     

Investigation of the role of diterpenes produced by marine pulmonatesTrimusculus reticulatus andT. conica

The intertidal pulmonate limpetTrimusculus reticulatus, which is found in caves or crevices along the California coast, was previously reported to contain two novel diterpenoids, 6-isovaleroxylabda-8,13-dien-7,15-diol (1) and 2,7-diacetoxy-6-isovaleroxylabda-8,13-dien-15-ol (2). Dissection of the animals prior to extraction revealed that the diterpenoids were concentrated in the mantle, foot, and mucus, but not in the viscera. The presence ofT. reticulatus or its mucus was toxic to veliger larvae of the sabellariid reef-building tube wormPhragmatopoma californica. The major diterpenoid1 was responsible for the observed larvicidal activity. Protection against overgrowth by settling invertebrate larvae is important for the survival of this sessile filter-feeding pulmonate. A related pulmonate,T. conica, which was found in similar intertidal habitats in New Zealand, contained 6-acetoxy-7-isovaleroxylabda-8,13-dien-15-oic acid (3) and 1,7,12,18-tetraacetoxy-cholest-5-en-3-ol (4). The diterpene3 was also localized in the foot, mantle, and mucus ofT. conica but was not larvicidal toP. californica.     

Learning of Herbivore-Induced and Nonspecific Plant Volatiles by a Parasitoid,Cotesia kariyai

Learning of host-induced plant volatiles by Cotesia kariyai females was examined with synthetic chemicals in a wind tunnel. Wasps were preconditioned by exposure to volatiles and feces simultaneously. A blend of four chemicals, geranyl acetate, -caryophyllene, (E)--farnesene, and indole, which are known to be specifically released from plants infested by host larvae Mythimna separata (host-induced blend), elicited a response in naive C. kariyai, but did not enhance the response after conditioning. A blend of five chemicals, (E)-2-hexenal, (Z)-3-hexen-1-ol, (Z)-3-hexen-1-yl acetate, -myrcene, and linalool, which are known to be released not only from plants infested by the host larvae, but also from artificially damaged plants or undamaged ones (unspecific blend), elicited little response in naive wasps, but significantly enhanced the wasps' response after conditioning. With a blend of the above nine chemicals, wasps could learn the blend at lower concentrations than they did in the nonspecific blend. Hence, both the host-induced and nonspecific volatile compounds appear to be important for C. kariyai females to learn the chemical cues in host location.     

Electrophysiological and behavioral responses of female Helicoverpa armigera to compounds identified in flowers of African marigold, Tagetes erecta

Seven electrophysiologically active compounds were detected in air-entrained headspace samples of live flowers of Tagetes erectaanalyzed by gas chromatography (GC) linked to a female Helicoverpa armigeraelectroantennograph (EAG) using polar and nonpolar capillary columns. These compounds were subsequently identified using GC linked to mass spectrometry as benzaldehyde, (S)-(–)-limonene, (R,S)-(±)-linalool, (E)-myroxide, (Z)--ocimene, phenylacetaldehyde, and (R)-(–)-piperitone. Electrophysiological activity was confirmed by EAG with a 1-g dose of each compound on filter paper eliciting EAG responses that were significantly greater than the solvent control response from female moths. Wind-tunnel bioassays with T. erectaheadspace samples, equivalent to 0.4 flower/hr emission from a live flower, elicited a significant increase in the number of upwind approaches from female H. armigerarelative to a solvent control. Similarly, a seven-component synthetic blend of EAG-active compounds identified from T. erectapresented in the same ratio (1.0 : 1.6 : 0.7 : 1.4 : 0.4 : 5.0 : 2.7, respectively) and concentration (7.2 g) as found in the natural sample elicited a significant increase in the number of upwind approaches relative to a solvent control during a 12-min bioassay that was equivalent to that elicited by the natural T. erectafloral volatiles.     

Identification of Semiochemicals Released During Aphid Feeding That Attract Parasitoid Aphidius ervi

Herbivore induced release of plant volatiles mediating the foraging behavior of the aphid parasitoid Aphidius ervi was investigated using the pea aphid, Acyrthosiphon pisum, feeding on broad bean, Vicia faba. Behavioral responses were studied using an olfactometer and a wind tunnel. Volatiles obtained by air entrainment of aphid infested plants were more attractive to A. ervi than those from uninfested plants, in both behavioral bioassays. GC-EAG of both extracts showed a number of peaks associated with responses by A. ervi, but with some differences between extracts. Compounds giving these peaks were tentatively identified by GC-MS and confirmed by comparison with authentic samples on GC, using two columns of different polarity. The activity of pure compounds was further investigated by EAG and wind tunnel assays. Results showed that, of the compounds tested, 6-methyl-5-hepten-2-one was the most attractive for A. ervi females, with linalool, (Z)-3-hexen-1-yl acetate, (E)--ocimene, (Z)-3-hexen-1-ol, and (E)--farnesene all eliciting significantly more oriented flight behavior than a solvent control. Foraging experience significantly increased parasitoid responses to these compounds, with the exception of (E)--farnesene. Time-course GC analysis showed that feeding of A. pisum on V. faba induced or increased the release of several compounds. Release of two of these compounds (6-methyl-5-hepten-2-one and geranic acid) was not induced by the nonhost black bean aphid, Aphis fabae. During the analysis period, production of (E)--ocimene remained constant, but 6-methyl-5-hepten-2-one, linalool, geranic acid, and (E)--farnesene appeared during the first day after A. pisum infestation and increased in concentration with increasing time of aphid feeding.     

Chemical and Chromatic Bases for Preferential Visiting By the Cabbage Butterfly, Pieris rapae, to Rape Flowers

Scent and coloration of corolla were examined as floral attributes responsible for preferential visiting by the cabbage butterfly, Pieris rapae, to rape flower, Brassica rapa. Floral volatile components that release the flower-visiting behavior of the butterfly were identified by chemical analyses, electroantennography (EAG), and two behavioral bioassays: proboscis extension reflex (PER) in response to odor and attraction to artificial flowers. GC and GC-MS analyses of the headspace volatiles from the flowers revealed the presence of six aromatic compounds, benzaldehyde, phenylacetaldehyde, benzyl alcohol, 2-phenylethanol, phenylacetonitrile, and indole in decreasing order of quantity. Of these, phenylacetaldehyde elicited the highest response in the PER assay. While benzyl alcohol, 2-phenylethanol, benzaldehyde, and phenylacetonitrile evoked moderate responses, the PER-eliciting activity of indole was very weak. In two-choice behavioral bioassays, artificial flowers scented with any one of these PER-active compounds attracted significantly more butterflies than control (unscented) flowers, whereas those treated with indole were almost inactive. The EAG activities of the six chemicals were not high and were about the same at a low dose (1 g), but phenylacetaldehyde elicited a much stronger response from both sexes at higher doses (10 and 100 g). An overall profile of EAG responses at a dose of 100 g was analogous to that of PER performance, suggesting that floral volatiles may be involved in close-range location or recognition of flowers rather than long-range attraction. By spectroscopic and UV-photographic examinations of rape flower, the central part of the corolla was found to absorb UV rays in marked contrast to the other parts, which reflected near-UV rays (_max = 350 nm). This indicates that the flower is endowed with a conspicuous nectar guide that is probably an important visual stimulus for attracting foraging adults of P. rapae. Consequently, the present findings strongly suggest that this elaborate pollination strategy of rape flower, characterized by its good combination of olfactory and visual attractiveness, accounts for preferential visiting by the cabbage butterfly to the flower.     

Inhibition of β-Glucosidase and Esterase by Tannins from Betula, Salix, and Pinus Species

The reaction of tannin fractions isolated from the bark of Betula, Salix, and Pinus species with two enzymes, -glucosidase and esterase, was investigated. The influence of precipitation to the hydrolytic capacity of -glucosidase also was studied. All tannins studied precipitated -glucosidase and esterase, and moderate differences in the precipitating capacities of the tannins were observed. Interestingly, complex formation between -glucosidase and tannin did not markedly affect the activity of the enzyme. Therefore, complex formation during the insect/herbivore feeding does not necessarily change the defense activity of phenolic glycosides or decrease activity of digestive enzymes.     

Floral Scent of Osmanthus fragrans Discourages Foraging Behavior of Cabbage Butterfly, Pieris rapae

Field observations revealed that flowers of Osmanthus fragrans attract few insect species despite their strong scent and vivid coloration (yellowish orange). Floral volatiles of the plant were thus examined for suspected repellency to Pieris rapae, a potential visitor that never visits this flower. The influence of odor components on the foraging behavior of adult butterflies was assessed by two behavioral bioassays: proboscis extension reflex (PER) in response to olfactory stimuli and the frequency of visits to artificial flowers. In the PER bioassays, a binary mixture composed of a test material and 2-phenylethanol (a positive standard compound) was used, and the negative effect of test materials was appraised based on the degree of suppression caused compared with the original PER performance evoked by 2-phenylethanol alone. Isopentane extract of the flower exhibited significant deterrency in the PER test and remarkable repellency in the flower-visiting experiment, thereby suggesting the presence of a repellent(s) in the floral volatiles. Fractionation of the extract by silica gel column chromatography yielded an active fraction (based on PER performance), which, upon GC and GC-MS analyses, was found to consist of -decalactone, -ionone, and linalool oxide (LO) isomers (furanosides and pyranosides) as major components together with small amounts of linalool and -ionone. -Decalactone was strongly deterrent in the PER test, and -ionone and all LO isomers were weakly deterrent, while the other compounds exhibited no deterrency. In the flower-visiting tests, -decalactone showed notable repellency, whereas -ionone had no repellent effect. Among the LO isomers, at least furanosides (cis/trans mixture) and cis-pyranoside were found to be weakly repellent. On the other hand, the butterfly showed strong to weak EAG responses to LO furanosides, cis-LO pyranoside, -decalactone, and -ionone in decreasing order of intensity. The present results clearly indicate that the repellency of a compound is correlated with its deterrent effect on PER but not necessarily with antennal sensitivity.     

Chemistry and Defensive Efficacy of Secretion of Burrowing Bug (Sehirus cinctus cinctus)

Adult Sehirus cinctus cinctus emit a volatile secretion from their metathoracic scent glands when tactually stimulated. We identified the volatile components by gas chromatography, high-performance liquid chromatography, and mass spectrometry. The secretion of both sexes contained (1R)-(+)--pinene, (1S)-(–)--pinene, -myrcene, (R)-(+)-limonene, and -terpinolene. Two additional compounds were found in only female secretions: (E)-2-hexenyl acetate and (E)-2-octenal. We also tested the defensive capability of this insect by offering it to various predators. Anoles, starlings, and a killdeer rejected S. c. cinctus after an initial sampling. These findings suggest that the secretion plays a defensive role.     

Female-produced oviposition deterrents of the cigarette beetle,Lasioderma serricorne (F.) (Coleoptera: Anobiidae)

Oviposition deterrents of the cigarette beetle,Lasioderma serricorne, were isolated from its adult body extract and found to be identical to (2S,3R,1'S)-2,3-dihydro-3,5-dimethyl-2-ethyl-6-(l-methyl-2-oxobutyl)-4H-pyran-4-one (-serricorone) and its lR-epimer (-serricorone) by spectroscopic evidence. Serricorone was previously found as one of the minor sex pheromone components of the same insect, and hence indicating bi-functional nature. The presence of two isomers in the body was proved by careful treatment. Each of them exhibited the same level of oviposition deterring activity, which was less potent than the crude body extract at an increased concentration.     

Response of the Seven-spot Ladybird to an Aphid Alarm Pheromone and an Alarm Pheromone Inhibitor is Mediated by Paired Olfactory Cells

Electrophysiological responses of adult seven-spot ladybirds, Coccinella septempunctata, to (E)--farnesene, an aphid alarm pheromone, and (–)--caryophyllene, a plant-derived alarm pheromone inhibitor, were investigated by recording from single olfactory cells (neurons) on the antenna. Cells having high specificity for each of the two compounds were identified. Furthermore, these two cell types were frequently found in close proximity, with a larger amplitude consistently recorded for the cell responding specifically to (E)--farnesene. Preliminary behavioral studies in a two-way olfactometer showed that walking adults were significantly attracted to (E)--farnesene; this activity was inhibited with increasing proportions of (–)--caryophyllene. The possible ecological significance of colocation or pairing of olfactory cells for semiochemicals with different behavioral roles is discussed.     

Spring migration of damson-hop aphid,Phorodon humuli (Homoptera,Aphididae), and summer host plant-derived semiochemicals released on feeding

Behavioral studies using an olfactometer demonstrated that spring migrants of the damson-hop aphid,Phorodon humuli, respond to semiochemicals released by spring migrants feeding on hop leaves. Samples of the total volatiles released on feeding were analyzed by coupled gas chromatographysingle cell recording techniques and showed the presence of several active components. Three compounds were identified, using coupled gas chromatography-mass spectrometry, as methyl salicylate, (E)-2-hexenal and-caryophyllene. These three compounds elicited responses from separate olfactory receptors on the antenna. In the olfactometer, both (E)-2-hexenal and-caryophyllene gave positive responses from spring migrants, and a mixture of the two compounds in the natural ratio was more attractive than (E)-2-hexenal alone. Addition of methyl salicylate eliminated the response to the active binary mixture.     

Correspondence of soldier defense secretion mixtures with cuticular hydrocarbon phenotypes for chemotaxonomy of the termite genus Reticulitermes in North America

Soldier defense secretions from samples of Reticulitermes collected in California, Nevada, Arizona, New Mexico, and Georgia were characterized and correlated with cuticular hydrocarbon phenotypes. Twenty-seven cuticular hydrocarbon phenotypes have been defined, and soldier defense secretion (SDS) phenotypes have been described for 25 of these. Forty-five terpenoid compounds were found, including monoterpenes, sesquiterpenes, and a few diterpenes. The monoterpenes include (–)--pinene, (–)--pinene, (–)-camphene, myrcene, (Z)- and (E)-ocimene, and (–)-limonene. The major sesquiterpenes produced are (+)--cadinene, (+)--cadinene aldehyde, (–)-germacrene A, germacrene B, -himachalene, and -bisabolene. Some SDS phenotypes pair with more than one cuticular hydrocarbon phenotype; however, with two exceptions, each hydrocarbon phenotype is associated with only one SDS phenotype. These chemical characterizations lend support to the conclusion that there are numerous undescribed species of Reticulitermes in North America.     

Selective Phytotoxic Activity of Withanolides from Iochroma australe to Crop and Weed Species

Phytotoxic effects of Iochroma australe extract and 4,7,20-oxowithanolides were examined in Petri dish bioassays at 10, 100, and 1000 ppm. The extract and the major constituent (17S,20R,22R)-4,7,20-trihydroxy-1oxowitha-2,5,24-trienolide (2) reduced growth of the radicle of the weeds Sorghum halepense (Monoct.) and Chenopodium album (Dicot.). Neither the extract nor withanolides had significant effect on germination or radicle length of the commercial crop species Lactuca sativa. From our experimental data we conclude that the withanolides tested here are natural products with selective herbicidal activity against weed species.     

Bioactive Steroids and Triterpenes from Melilotus messanensis and Their Allelopathic Potential

The aerial parts of Melilotus messanensis (sweet clover) afforded, from moderately and polar bioactive fractions, 11 triterpenes and five steroids. A series of aqueous solutions at 10^–4–10^–9 M of five steroids and nine triterpenes was monitored to test their effects on germination and growth of the monocots Hordeum vulgare and Allium cepa, and the dicots Lactuca sativa (var. nigra and var. romana) and Lycopersicon esculentum. An important stimulatory effect on H. vulgare germination (between 40% and 80% for almost all tested compounds) was observed. Some considerations about the ecological role of triterpenes on M. messanensis are made. In addition to known lupane triterpenes (platanic acid and 3,28,30-lup-20(29)-enetriol), oleanane triterpenes (soyasapogenol B, soyasapogenol G, and messagenolide), a gammacerane triterpene (gammacer-16-en-3-one), five new lupane triterpenes (messagenic acids A–E: (27-cis-4-hydroxycinnamoyloxy)-betulinic acid; 27-(trans-4-hydroxycinnamoyloxy)betulinic acid; 20(S)-3-hydroxy-29-oxolupan-28-oic acid; 3,30-dihydroxylup-20(29)-en-28-oic acid; and 3,20-dihydroxylup-18(19)-en-28-oic acid, respectively), and sterols (-sitosterol, ergosterol peroxide, 7-hydroxysitosterol, 7-hydroxysitosterol, and 7-oxositosterol) were isolated and chemically characterized. Their structures and stereochemistry were elucidated by spectroscopic methods (one- and two-dimensional ¹H and ¹³C NMR, IR, MS).