Attractancy toOryzaephilus surinamensis (L.) of volatile materials isolated from vacuum distillate of heat-treated carobs

Vacuum distillation of heat-treated carobs gave an aqueous, colorless, sweet-smelling distillate which was tested over a wide range of concentrations and found to be highly attractive to adultOryzaephilus surinamensis (L.). The materials responsible for the aroma were isolated from the distillate by saturating with sodium chloride and extracting into diethyl ether as separate acidic, neutral, and basic fractions. The extraction efficiency was checked by recombining portions of the three fractions and replacing the diethyl ether with water to give a reconstituted distillate; this was almost as attractive as the original distillate. Bioassay of aqueous solutions of the three separate fractions showed that the acidic was attractive, while the neutral and basic had little effect. The five major components of the acidic fraction were found to be acetic, isobutyric,n-butyric, 2-methylbutyric, and hexanoic (caproic) acids. Bioassay of these in aqueous solution, both separately and combined, showed that hexanoic acid was the most attractive and may be responsible for both the longer-lasting attractive effect of the carob distillate and for the effectiveness of carobs themselves used in bait bags to detect stored product insects.     

Biocidal activity of algal toxins against immature mosquitoes

Extracts of two species of green algae, filamentousRhizoclonium hieroglyphicum Kütz and a phytoplankton,Chlorella ellipsoidea Gerneck, obtained with solvents in the laboratory were assayed againstAedes aegypti L.,Culex quinquefasciatus Say, andCuliseta incidens (Thomson). On extraction with petroleum ether, groundR. hieroglyphicum yielded an active crude extract which was chromatographed on a neutral alumina column and eluted consecutively with petroleum ether, benzene, and methanol. All three eluted fractions were found to induce significant mortality in test mosquito species. The benzene-eluted fraction was the least toxic. The methanol-eluted fraction was the most toxic to all species and exhibited juvenile hormone-like activity; it also caused morphogenetic changes in emerging adults. All three fractions delayed the rate of development of mosquito larvae by 2–5 days. Three supernatants ofC. ellipsoidea obtained on different occasions were tested against first instars ofC. quinquefasciatus. After the confirmation of their activity, all supernatants were extracted with diethyl ether, combined, and assayed against first- and fourth-stage larvae of the three mosquito species. The first-stage larvae ofC. quinquefasciatus andC. incidens were approximately three times more suspectible than those ofA. aegypti. However, fourth-stage larvae of the former two species were about twice as susceptible to the extract as those of the latter species. Dead first-stage larvae of all the species had a shrunken appearance. In general,C. ellipsoidea extract was quicker acting than that ofR. hieroglyphicum.     

Patterns of Oviposition Stimulants for Carrot Fly in Leaves of Various Host Plants

Undamaged leaves of 12 host-plant species differing widely in acceptability to ovipositing carrot flies were extracted with a microwave-assisted method with hexane as solvent. The highly stimulatory diethyl ether fraction obtained by separation on a silica gel column was semiquantitatively analyzed by GC-MS for previously identified oviposition stimulants of the carrot fly (phenylpropenes, fluranocoumarins, polyacetylenes). Various plant species exhibited widely differing profiles of these compounds. In choice assays, moderate numbers of eggs were deposited underneath surrogate leaves sprayed with fractions that contained high amounts of just one type of compound and low amounts of the other two types. Only fractions with medium to high levels of at least two compound classes elicited strong ovipositional responses (e.g., phenylpropenes and polyacetylenes in Daucus carota, furanocoumarins and polyacetylenes in Heracleum sphondylium and Conium maculatum). None of the examined plants contained high quantities of all three compound classes. The contents of the stimulants seemed to account in a synergistic manner for the variation in activity of the diethyl ether fraction. However, they could not explain adequately the observed preference hierarchy of the carrot fly for the host-plant species.     

Extracts of Flourensia cernua Reduce Consumption of Alfalfa Pellets by Sheep

Effects of three extracts (hexanes, ether, and ethanol) from tarbush (Flourensia cernua) on intake of alfalfa pellets by lambs were examined. Forty-five ewe lambs were fed one of five treatments for five days (randomized complete block, three lambs per block on each treatment). Treatments were alfalfa pellets (CON) or alfalfa pellets plus ethanol carrier (CAR), hexanes extract (HEX), ether extract (ETH), or ethanol extract (ETOH). Extracts were applied to alfalfa pellets at the same concentration as in an equivalent amount of tarbush (as fed basis) in experiment 1 and at 10-fold dilutions of that concentration in experiment 2. Treatments were isolated from tarbush leaves by using a sequential extraction with hexanes, diethyl ether, and 100% ethanol. Lambs received 640 g of alfalfa pellets (dry matter basis) each morning and intake was monitored during a 20-min interval. Lambs were maintained and fed alfalfa pellets (4.7% of body weight) as one group except during this interval. In experiment 1, mean intake by lambs during the 20-min interval was 361, 393, 204, 212, and 228 g for CON, CAR, HEX, ETH, and ETOH, respectively (SEM = 28.9). All three extracts decreased intake (P < 0.001) compared to CON or CAR. Intake did not differ among the three extracts (HEX, ETH, and ETOH) or between the two controls (CON and CAR). Mean intake did not differ among treatments in experiment 2 (468, 455, 389, 381, and 431 g for CON, CAR, HEX, ETH, and ETOH, respectively; SEM = 30.5; P = 0.187). Several compounds are probably responsible for the low palatability and differential use of tarbush typically exhibited by livestock.     

Isolation and identification of (3-methoxyphenyl)acetonitrile as a phytotoxin from meadowfoam (Limnanthes alba) seedmeal

Ethyl ether, ethanol, and water extracts of meadowfoam (Limnanthes alba Hartweg ex. Benth.) seedmeal were prepared and bioassayed against velvetleaf (Abutilon theophrasti Medicus) and wheat (Triticum aestivum L. Cardinal). Both the ethyl ether and ethanol fractions, but not the water extract, inhibited velvetleaf and wheat radicle elongation. Fractionation of the extracts indicated that (3-methoxyphenyl)acetonitrile (3-MPAN) was the active compound from both extracts, comprising >97% of the active ethanol fraction. 3-Methoxybenzyl isothiocyanate, which had been previously shown to be the major breakdown product of glucolimnanthin, the majorL. alba glucosinolate, was not detected in either extract. Radicle elongation of velvetleaf and wheat were inhibited by 3-MPAN with I₅₀ (the concentration required to inhibit growth by 50%) values of approximately 4 × 10^–4 M (velvetleaf) and 7×10^–4 M (wheat).Mention of firms or products does not imply endorsement by the U.S. Department of Agriculture over other firms or products not mentioned.     

Chemical inhibition of fire-prone grasses by fire-sensitive shrub,Conradina canescens

In an investigation of potential chemical activity of fire-sensitive shrubs in Florida's sand pine scrub community, bioassays of foliar washes ofConradina canescens showed significant inhibitory activity on three native grasses that are known to fuel frequent surface fires; inhibition was concentrated seasonally in spring and summer. Application of runoff fromConradina leaves to one of the grasses caused a 50% reduction in growth over a 20-week period. Isolation of the biologically active fractions from the fresh leaves ofC. canescens yielded numerous monoterpenes, a number of which were identified from a GC-MS reference library and/or MS comparison to authentic compounds: 11 from the diethyl ether extract, 11 from steam distillation, and four from the foliar leaf wash. Numerous other monoterpenes present in the extractions were unknown. The terpenoid fraction completely inhibited seed germination of one of the native grasses and of lettuce. Saturated aqueous solutions of nine of the monoterpenes inhibited germination and radicle growth of two native grasses. SEM views of the leaf surfaces ofConradina reveal secretory trichomes that appear to be the source of the monoterpenes as well as the triterpene, ursolic acid. The biological activity ofC. canescens as a fire-sensitive component of the scrub community is reviewed in light of the chemical evidence.     

Allelopathic potential of sorghum-sudangrass hybrid (sudex)

Experiments were conducted under controlled conditions to investigate the apparent allelopathic effects of sudex [Sorghum bicolor (L.) Moench ×Sorghum Sudanese (P.) Stapf, cv. FFR 201] on weed and vegetable species. Allelopathic potential, as measured by radicle elongation of herbaceous indicator species, decreased with increasing sudex age. Greatest potential allelopathic activity of sudex shoot tissue was observed when sudex was collected at 7 days of age. Small-seeded broadleaf species were more inhibited in the presence of sudex shoot tissue than were grass species. Two major phytoinhibitors were isolated from aqueous extracts of sudex shoot material by partitioning with diethyl ether, followed by thin-layer and liquid column chromatography. Phytoinhibitors were identified asp-hydroxybenzoic acid andp-hydroxybenzaldehyde, potentially the enzymatic breakdown products of the cyanogenic glycoside dhurrin. The I₅₀ values of these compounds using a cress (Lepidium sativum L.) seed bioassay were 140 and 113 g/ml for the acid and aldehyde, respectively. Sudex tissue collected at 7 days of age possessed a greater percentage of these phytoinhibitors on a per gram basis than did older sudex tissue. As sudex tissue age increased, the percentage ofp-hydroxybenzaldehyde in ether extracts of tissue also increased, while the percentage ofp-hydroxybenzoic acid decreased.Journal article No. 88-10-163 of the Kentucky Agricultural Experiment Station.     

Cardenolides fromErysimum cheiranthoides: Feeding deterrents toPieris rapae larvae

Larvae of the cabbage butterfly,Pieris rapae, refuse to feed on the wild mustard,Erysimum cheiranthoides, due to the presence of alcoholextractable deterrents. The active components were extracted inton-BuOH, and this extract was separated into four fractions (I–IV) by reverse-phase HPLC. Fractions III and IV retained the feeding deterrent activity. The activity of fraction III was found to be due to the cardenolide diglycosides 1 and 2, which were previously reported as oviposition deterrents for gravidP. rapae butterflies. Three active compounds were isolated from fraction IV by column chromatography on silica gel followed by reverse-phase HPLC. These compounds were identified as a monoglycoside, digitoxigenin 3-O--D-glucoside (4), and two diglycosides, glucodigigulomethyloside (5) and glucodigifucoside (6). An additional cardenolide isolated from fraction II was identified as cheirotoxin (7). All compounds were identified by UV, NMR (¹H and¹³C), and mass spectrometry, as well as hydrolysis experiments. The feeding deterrent activity of these compounds was compared with that of related commercially available chemicals and other compounds isolated fromE. cheiranthoides.     

Excess molar enthalpies for the binary and ternary mixtures of ether compounds (di-isopropyl ether, di-butyl ether, propyl vinyl ether) with ethanol and isooctane at 298.15 K

Excess molar enthalpy (H ^E ) data at 298.15 K are reported for the binary systems di-isopropyl ether (1)+ethanol (2), di-isopropyl ether (1)+isooctane (2), ethanol (1)+isooctane (2), di-butyl ether (1)+ethanol (2), di-butyl ether (1)+isooctane (2), propyl vinyl ether (1)+ethanol (2) and propyl vinyl ether (1)+isooctane (2). These data were obtained by using an isothermal flow calorimeter. The experimental binary H ^E data were well correlated with the Redlich-Kister model, and infinitely dilute partial excess molar enthalpies for each binary were calculated with the fitted Redlich-Kister parameters. Additionally, the isoclines of H ^E for ternary systems di-isopropyl ether (1)+ethanol (2)+isooctane (3), di-butyl ether (1)+ethanol (2)+isooctane (3) and propyl vinyl ether (1)+ethanol (2)+isooctane (3) at 298.15 K were calculated by using the Radojkovič equation. H ^E for all the measured systems in this work shows that mixing is endothermic.     

Vulgarone B,the antifungal constituent in the steam-distilled fraction of Artemisia douglasiana

Antifungal activity of the steam distilled essential oil fraction of Artemisia douglasiana was detected by bioautography on silica gel TLC plates against three Colletotrichum spp. The active principle was isolated by bioassay-directed fractionation using column chromatography followed by crystallization and was characterized as vulgarone B by ¹H and ¹³C NMR and GC-MS. Antifungal activity of vulgarone B was further evaluated using 96-well microtiter assay against Colletotrichum acutatum, C. fragariae, C. gloeosporioides, and Botrytis cinerea. In addition, the antifungal activity of vulgarone B and verbenone, and their corresponding alcohols was tested by bioautography and microtiter assay. Structure–activity studies revealed that the , -unsaturated carbonyl functionality is a prerequisite for the antifungal activity of these mono and sesquiterpene ketones. This is the first report of antifungal activity of vulgarone B. The yield of essential oil from A. douglasiana is about 0.6–0.8% by weight of the dry material, including plant stems.     

Himachalol andβ-himachalene: Insecticidal principles of himalayan cedarwood oil

Chromatographic fractions of Himalayan cedarwood oil (Cedrus deodara) were bioassayed against the pulse beetle (Callosobruchus analis F.) and the housefly (Musca domestica L.). Almost all fractions showed insecticidal activity against both test species. Fractions I and V led to the highest mortality and also produced a quick knockdown effect. Fractions I and V, after rechromatography and purification, yielded himachalol (3%) and -himachalene (31%), based on essential oil weight, respectively. Further evaluation of these two naturally occurring sesquiterpenes indicated 97.5% mortality at 0.56 mol/insect against the pulse beetle. These biologically active natural products of plant origin may serve as a suitable prototypes for development of commercial insecticides.CIMAP Publication No. 598.     

Antifungal activity of volatile fractions of essential oils from four aromatic wild plants in israel

Essential oils were extracted fromMajorana syriaca, Satureja thymbra, Micromeria fruticosa, andSalvia triloba, and their volatile fractions were tested for their antifungal activity against the soil-borne pathogensFusarium oxysporum andMacrophomina phaseolina and the foliar pathogensBotrytis cinerea andExserohilum turcicum. Results showed a fungistatic effect of 1, 2.5, and 5l of the various essential oils on fungal mycelium growth. The most significant effect was exerted by essential oils extracted fromM. syriaca, which inhibited the growth ofB. cinerea by 44% and of all the other fungi tested by 100%.Contribution from the Agricultural Research Organization, the Volcani Center, Bet Dagan, Israel. No. 3555-E, 1992 series.     

Sex pheromones of male yellowfin Baikal sculpin (Cottocomephorus grewingki): Isolation and chemical studies

Three components of the male yellowfin Baikal sculpin pheromonal signal have been isolated from urine by diethyl ether extraction, thinayer chromatography (TLC), and high-performance liquid chromatography (HPLC). Using mass spectrometry, we have identified two of them as testosterone (T) and 11-hydroxytestosterone (11HT). These steroids are synthesized in testes during full spermatogenesis, and they are excreted in male urine with milt. The third component is not a steroid. It is more likely to be a polyene alcohol (farnesol). 2Z,6E-Farnesol possesses behavioral activity.     

Isolation and identification of house fly,Musca domestica L., repellents from pepper tree,Schinus molle L.

Foliage from the pepper tree,Schinus molle L., is traditionally used in Ethiopia to repel house flies,Musca domestica L. The volatile extracts of pepper tree leaves were shown to have repellent and feeding-deterrent activity against house flies in a two-choice laboratory bioassay. High-performance liquid chromatographic fractionation of steam-distilled volatiles from leaves, monitored by laboratory bioassays, demonstrated that bioactivity is associated with two compounds,cis-menth-2-en-1-ol andtrans-piperitol. The absolute configuration of the latter was established as (1S,6S)-piperitol by comparison of acetyl lactate derivatives. Racemic compounds were synthesized from piperitone, and bioassays with house flies indicatedtrans-piperitol to be the most active house fly repellent.     

Oviposition stimulants and deterrents regulating differential acceptance ofIberis amara byPieris rapae andP. napi oleracea

Iberis amara (Cruciferae) contains both stimulants and deterrents that are involved in regulating oviposition byPieris rapae andP. napi oleracea. The most active deterrents toP. rapae isolated from butanol extracts of the plant were found to be 2-O--d-glucosyl cucurbitacin I and 2-O--d-glucosyl cucurbitacin E. However,P. napi oleracea was behaviorally insensitive to these compounds and was only weakly deterred by other individual fractions of the butanol extract. Stimulant activity of the postbutanol water extract ofI. amara was associated with glucosinolates. The most abundant of these was identified as sinigrin, and a relatively minor component was shown to be glucoiberin. The isolated sinigrin was more stimulatory toP. rapae than was the glucoiberin-containing fraction, butP. napi oleracea was stimulated as strongly by the glucoiberin fraction, even though the concentration of this compound was much lower. The contrasting responses of the twoPieris species to the deterrents and stimulants inI. amara can explain the differential acceptance of the plant by these butterflies.     

Seed germination and growth inhibitory cadinenes fromEupatorium adenophorum spreng

Chloroform extract of the aerial parts ofEupatorium adenophorum Spreng was fractionated and examined for growth inhibition. Bioassay-directed fractions of the plant materials afforded three known cadinenes and -sitosterol. The effects of different fractions as well as isolated cadinenes were determined usingAllium cepa, Raphanus sativus, andCucumis sativus seeds. Three-day exposure to these cadinenes significantly inhibited germination and seedling growth of all three assay seeds. The degree of inhibition was dependent upon seed species and the concentrations of the compounds tested. Cadinene(1) was found to be more inhibitory to the seeds tested and the activity of the cadinene(3) was less than that of(1) and(4).     

Isolation of the diterpenoids, ent-kauran-16alpha-ol and ent-atisan-16alpha-ol, from sunflowers, as oviposition stimulants for the banded sunflower moth, Cochylis hospes

Two diterpenoid alcohols, ent-kauran-16-ol (1) and ent-atisan-16-ol (2), were isolated from pre bloom (R3-R4 stage) sunflower heads as oviposition stimulants for the banded sunflower moth, Cochylis hospes. Fractionation of a sunflower head extract, by normal-phase flash column chromatography, resulted in an early eluting fraction exhibiting significant activity in an egg-laying bioassay. Compounds 1 and 2, along with ent-trachyloban- 19-oic acid (3) and ent-kaur-16-en-19-oic acid (4), were isolated as the major components of this fraction and identified by their NMR and mass spectra. The purified compounds were individually tested for ovipositional activity in dose-response bioassays. In these bioassays, compounds 1 and 2 gave linear dose responses, with increasing numbers of eggs laid as the dosage of either increased. Compounds 3 and 4 failed to stimulate significant egg-laying at any of the dosages tested. A factorial design bioassay, using compounds 1 and 2, showed that 1 was relatively more stimulatory than 2, and that there was no synergistic effect on oviposition when the two compounds were combined.     

Phytotoxicity of Vulpia Residues: II. Separation, Identification, and Quantitation of Allelochemicals from Vulpia myuros

Phytotoxicity-based extraction and fractionation were employed to separate and purify the allelochemicals from an aqueous extract of vulpia (Vulpia myuros) residues. Further analyses, identification, and quantitation of these allelochemicals were conducted by gas chromatography–mass spectrometry on the most toxic fraction. Twenty-one compounds were identified in the ether fraction from the vulpia residue aqueous extract. They were catechol, hydroquinone, 3,4-dimethoxyphenol, pyrogallol, coniferyl alcohol, and benzoic, succinic, hydrocinnamic, salicylic, protocatechuic, vanillic, gentisic, syringic, p-hydroxybenzoic, -hydroxybenzenepropanoic, p-hydroxyphenylacetic, p-hydroxybenzenepropanoic, hydroferulic, p-coumaric, hydrocaffeic, and ferulic acids. A chromatographic internal standard method with multiple-point calibration graphs was used to quantify the identified compounds. Quantities in the vulpia residues ranged from 1.36 to 81.0 g/g dry residue, in total accounting for 0.05% of the dry weight residue. The combined syringic and hydroferulic acids were present in the largest amount, 140.11 g/g residue. Vanillic, succinic, p-hydroxybenzenepropanoic, and salicylic acids were next in amount, ranging from 37.24 to 81.24 g/g residue. Catechol, hydrocinnamic acid, and hydroquinone were present in the smallest quantities, ranging from 1.36 to 1.82 g/g residue. The remainder of the compounds were intermediate in amount, ranging from 2.33 to 18.1 g/g residue.     

Response on the ground of bark beetle and weevil species colonizing conifer stumps and roots to terpenes and ethanol

Responses of threeHylastes species,Dryocoetes autographus, and twoHylobius species to terpenes and ethanol were studied in field experiments on clear-cut forest sites in Sweden using baited ground traps.-Pinene alone did not attract any of the six species. A terpene blend (spruce turpentine consisting mainly of-pinene,-pinene, and 3-carene) attractedHylastes cunicularius, H. brunneus, andHylobius abietis in some experiments, but not in others. The attractiveness of ethanol also varied; the only species consistently attracted wasH. abietis. Baits containing both terpenes and ethanol, particularly the combination of spruce turpentine and ethanol, were attractive to all species exceptHylobius pinastri. InH. abietis, the terpene plus ethanol/ ethanol catch ratios increased during early summer. Seasonal differences in catch levels were observed inH. cunicularius andH. abietis. The addition of-pinene reduced the attractiveness of the combination of spruce turpentine and ethanol toH. cunicularius, H. opacus, andD. autographus. The differences in response to the volatiles between species are probably related to differences in reproductive behavior and host preferences.     

Response ofAedes triseriatus larvae to fatty acids ofCladophora

The activity of a methanol extract residue fromCladophora glomerata against larvae ofAedes triseriatus is confirmed. Fractionation of this residue by solubility in solvents of varying polarity is monitored by bioassay withA. triseriatus larvae. The presence of fatty acids in the active hexane soluble fraction and saponified material obtained therefrom is indicated by¹H NMR and IR. The identification of the individual fatty acids is achieved by co-injection and mass GLC of their methyl esters. Saturated and unsaturated fatty acids are screened against larvae and the LD₅₀ values of the most active acids are determined. The LD₅₀ of capric [100], lauric [120], myristic [140] and palmitoleic [161 (9c)] are 14,7, 4 and 3ppm respectively. The most active acids 120, 140 and 161 (9c) are present in all the fractions examined and are released to water by the powdered alga at pH 8.5.