Chemotactic tube-foot responses of a spongivorous sea starPerknaster fuscus to organic extracts from antarctic sponges

Hexane, chloroform, and methanol extracts of 18 species of antarctic sponges were tested for their ability to induce sustained tube-foot retraction in the antarctic spongivorous sea starPerknaster fuscus. Extracts were imbedded in silicone and used to coat the tip of a glass rod, which was allowed to contact an extended tube-foot. Retraction times were measured and compared with three controls: contact with a glass rod coated with a hexane extract of fish (feeding stimulant), contact with the glass rod alone (mechanical control), and contact with the glass rod coated with silicone alone (silicone control). Only extracts of the spongeMycale acerata did not elicit significantly longer tube-foot retraction times than controls for at least one of the three organic extracts. Hexane sponge extracts elicited the lowest levels of significant tube-foot responses, with only 39% of the sponge species tested showing activity in this fraction. In contrast, chloroform and methanol extracts elicited a significant tube-foot retraction response in 73% and 78% of the species tested, respectively. This indicates that in this assay repellent metabolites are generally more polar substances. It remains to be determined that secondary metabolites are responsible for all of the tube-foot retraction responses detected in sea stars exposed to sponge extracts; bioactive secondary metabolites have been isolated from a number of these antarctic sponges. It may be of ecological significance that the two rapidly growing sponges,Homaxinella balfourensis andMycale acerata, were either not repellent or had low repellency, and thatM. acerata is the primary dietary item ofPerknaster fuscus.     

Chemistry and chemical ecology of the Bahamian spongeAplysilla glacialis

Chemical investigation of the secondary metabolites of the marine spongeAplysilla glacialis collected at French Wells, Crooked Island, Bahamas, resulted in the isolation and characterization of four diterpenes, two sterol endoperoxides, and two methylated primary metabolites. Some of these compounds, along with crude extracts of the sponge, were investigated for their ability to deter fish predation, affect the fouling of surfaces, and inhibit the growth of marine microorganisms. The diterpene manoöl (3), cholesterol endoperoxide (4), and the crude nonpolar extract ofA. glacialis were shown to deter feeding by a natural assemblage of fish predators in an in situ assay conducted at French Wells. Pure secondary metabolites and crude extracts ofA. glacialis also were tested in a laboratory fish feeding assay employing the wrasseThalassoma lunare. A mixture of sterol endoperoxides was isolated from the mucus that coats the surface ofA. glacialis and is exuded in large quantities when the sponge is disturbed. These compounds are thereby distributed in a manner in which they can best serve a defensive role for the sponge. An in situ assay was designed to determine the effect that pure secondary metabolites and crude extracts have on the fouling of surfaces. Manoöl (3) and cholesterol endoperoxide (4) were determined to increase the rate of fouling when compared to control surfaces. 1-Methyladenine (5) was identified as an antimicrobial constituent ofA. glacialis that inhibited the growth of four marine bacteria isolated from seawater samples collected at French Wells.This work was presented at the Gordon Research Conferences (Marine Natural Products Chemistry), February 1990, Oxnard, California.     

Feeding behavior of graminivorous grasshoppers in response to host-plant extracts,alkaloids, and tannins

Secondary metabolites exhibit the potential to direct food selection by grass-feeding (graminivorous) grasshoppers. We examined the effects of plant extracts and representative secondary metabolites on the feeding behavior of two such grasshoppers,Ageneotettix deorum (Scudder) andPhoetaliotes nebrascensis (Scudder). Three alkaloids and two tannins were bioassayed for their activity as feeding deterrent allelochemicals, as were extracts from the foliage of the graminoids commonly eaten by these grasshoppers:Agropyron smithii Rydb.,Andropogon hallii Hack.,Andropogon scoparius Michx.,Bouteloua gracilis (H. B. K) Lag. ex Griffiths,Carex heliophila Mack. andStipa comata Trin. & Rupr. Alkaloids strongly deterred feeding but tannins only exhibited a weak effect, even when present at four times the concentration of total phenolics typical for these graminoids. Host-plant extracts also exhibited weak effects, such that we found no evidence for either strong deterrence or phagostimulation. Our results for alkaloids and host-plant extracts are consistent with the view that grass-feeding grasshoppers may be restricted to graminoids because of: (1) the presence of deterrents in nonhosts and (2) the absence of deterrents in hosts. However, our data for tannins show that these are unlikely to be effective barriers to herbivory by these grasshoppers.     

Unpalatable Compounds in the Marine Gastropod Dolabella auricularia: Distribution and Effect of Diet

Sea hares are a rich source of novel secondary metabolites, most of which are derived from their algal diet, but the natural function(s) of these metabolites are largely unknown. We used field and laboratory assays to measure the palatability of extracts from the tissues, ink, and eggs of Dolabella auricularia. Digestive-gland extracts contained a wide variety of secondary metabolites, including the red algal compound prepacifinol epoxide and its derivative johnstonol, and they were unpalatable to reef fishes. Skin extracts were moderately unpalatable, but our bioassay-guided fractionation led us to (–)-7-dehydrocholesterol, rather than to an algal secondary metabolite. Ink extracts were consistently unpalatable to reef fishes only at high concentrations, suggesting either that ink must be concentrated to deter predators, that unpalatable components of ink rapidly decompose, or that ink has other functions. Unpalatability of ink was traced to a purple fraction, consistent with the hypothesis that the active compound is aplysioviolin, a known ink constituent modified from a red algal pigment. Egg extracts were moderately unpalatable; however, we could not trace this activity to any algal-derived secondary metabolite. Body-wall extract was highly palatable. Our results suggest that dietary-derived secondary metabolites play a role in chemical defense of D. auricularia via the ink, but are not responsible for unpalatability of skin or eggs. Accumulation of dietary-derived metabolites in the digestive gland may occur to detoxify a chemically rich diet, rather than or in addition to deterring predators.     

The role of primary and secondary metabolites in chrysanthemum resistance toFrankliniella occidentalis

The presence of deleterious secondary metabolites to western flower thrips (WFT) (Frankliniella occidentalis Pergande) in several chrysanthemum (Dendranthema grandiflora Tzvelev) cultivars was tested. WFT performance on leaf sap mixed with artificial diet was compared to WFT performance on a control of artificial diet mixed with water. Leaf sap of six of 10 chrysanthemum cultivars tested had a significant negative effect on WFT performance and therefore contained deleterious secondary metabolites. Leaf sap of a resistant and a susceptible chrysanthemum cultivar were fractionated by gel to study the role of secondary metabolites in greater detail. None of the 20 individual fractions obtained had a negative effect on WFT performance, whereas after mixing of these fractions activity was found again. Moreover, WFT performance on the fraction mixture of the resistant cultivar was significantly lower than on the susceptible fraction mixture. The results suggest that WFT resistance in the cultivars studied was caused by several secondary metabolites that had an additional or synergistic negative effect on WFT performance. The role of a deficiency of primary metabolites in WFT resistance of chrysanthemum was also investigated. WFT performance on leaf sap with and without extra primary metabolites (artificial diet) was compared. The variance in WFT performance was significantly smaller when extra primary metabolites were added; WFT performance on leaf sap with and without extra primary metabolites was not correlated. The results suggest that resistance was partly caused by a deficiency of primary metabolites.     

Effects of secondary metabolites from marine algae on feeding by the sea urchin,Lytechinus variegatus

A bioassay was developed to test the hypothesis that secondary metabolites from marine algae affect feeding by sea urchins. During experiments measuring chemoreception and gustation, feeding by the regular sea urchin,Lytechinus variegatus (Lamarck), was inhibited by extracts from the green marine alga,Caulerpa prolifera (Forsskal) Lamouroux andCymopolia barbata (Linneaus) Lamouroux. Cymopol, a monoterpene-bromohydroquinone component ofC. barbata, was partially responsible for the inhibited feeding observed in tests of theCymopolia crude extract. Caulerpenyne, an oxygenated sesquiterpene fromC. prolifera, was responsible for all of the urchin feeding inhibition observed in tests of theC. prolifera crude extract. Feeding was not affected by: (1) extracts from several otherCaulerpa species includingC. mexicana (Sonder) J. Agardh,C. ashmaedii Harvey,C. racemosa v.macrophysa (Kutzing) Taylor, andC. racemosa v.laetevirens (Montagne) Weber-van Bosse; (2) caulerpin, an indole-containing pigment isolated from all of theCaulerpa species exceptC. mexicana; and (3) an extract from the red marine alga,Gracilaria foliifera v.angustissima (Harvey) Taylor, which has no known secondary metabolites. Feeding inhibition was independent of the test diameter which correlated with the reproductive state of the urchins. Feeding inhibition was also independent of the starvation periods between experiments, and the temperature and salinity in ranges tolerated byL. variegatus obtained from the Florida Gulf Coast. The data strongly suggest that at least one alga,Caulerpa prolifera, is chemically defended against a dominant omnivore in its community.     

Dynamics of Algal Secondary Metabolites in Two Species of Sea Hare

The function of acquired algal secondary metabolites in sea hares is the subject of debate, in part because the dynamics/processing of metabolites by sea hares is poorly understood. This study investigates the dynamics of red algal secondary metabolites in two sea hares, Aplysia parvula and Aplysia dactylomela. Secondary metabolite levels were quantified for the dietary red algae Laurencia obtusa and Delisea pulchra and for sea hares collected from these seaweeds in the field. The patterns and dynamics of algal secondary metabolites were further investigated in the laboratory by quantitative analysis of secondary metabolites in sea hares grown on diets of L. obtusa, D. pulchra, or the green alga Ulva sp. Sea hares accumulated the most abundant metabolites from each red alga, the terpene palisadin A from L. obtusa, and the halogenated furanone 3 from D. pulchra, and stored a greater proportion of these metabolites than other algal metabolites. A. parvula accumulated D. pulchra metabolites at much higher levels than L. obtusa metabolites. A. dactylomela accumulated similar concentrations of L. obtusa metabolites to A. parvula. The loss of L. obtusa metabolites by A. dactylomela matched that expected for dilution of metabolites via growth of the sea hares. However, the loss of L. obtusa metabolites by A. parvula was faster than predicted for growth alone, suggesting that metabolites were actively metabolized or excreted. Data for the loss of D. pulchra metabolites by A. parvula was equivocal. The secretions of A. parvula fed D. pulchra or L. obtusa were analyzed for the presence of algal secondary metabolites to investigate one possible path of excretion. L. obtusa secondary metabolites were detected in the mucous and opaline secretions of A. parvula, but D. pulchra metabolites were not detected in any secretions. The deployment of L. obtusa secondary metabolites in secretions by A. parvula may explain the more rapid rate of loss of these compounds and is consistent with a possible defensive role for acquired metabolites.     

Evidence for chemical defense in tropical green algaCaulerpa ashmeadii (Caulerpaceae: Chlorophyta): Isolation of new bioactive sesquiterpenoids

Results of field feeding preference studies with 12 species of tropical green algae of the genusCaulerpa showed thatC. ashmeadii was preferred least by herbivorous fishes. Chemical investigations ofC. ashmeadii demonstrated the presence of high concentrations of sesquiterpenoid metabolites. The chemical isolation and structural elucidation of five majorC. ashmeadii metabolites, as well as the results of field feeding preference, antimicrobial, and ichthyotoxicity assays demonstrating the biological activities of these metabolites are reported here.     

Attraction of predatorily naive postlarval lobsters to extracts of metabolites of common prey:Mytilus edulis,Mya arenaria,Cancer irroratus,andAsterias vulgaris

Postlarval lobsters (4th–7th stage) exclusively fed frozen brine shrimp (Artemia saline) were assayed for food-search response to extracts and metabolites from four common prey: soft clams (Mya arenaria), blue mussels (Mytilus edulis), rock crabs (Cancer irroratus), and sea stars (Asterias vulgaris). Concentrations of soluble primary amines, protein, and ammonia in prey tissues and metabolites were determined. No significant responses were observed for any prey metabolites diluted to 1 and 10%, while onlyA. vulgaris evoked a significant response at full strength, suggesting that predatorily naive lobsters have yet to develop more pronounced chemosensory responses shown by field-collected lobsters. Removal of protein with retention of small-molecular-weight polar molecules did not appear to affect response to prey extracts. EC₅₀s, as micromoles per liter amines, computed from prey extract dose-response curves indicate differences per unit amine between prey extracts, withA. vulgaris extract more potent as an attractant than either bivalve extract.C. irroratus extract was equally attractive as the other three extracts. Ammonia levels excreted into seawater over 3 hr were similar for all prey species, while soluble primary amines and proteins were undetectable. Ammonia and protein per gram whole prey varied significantly between extracts of prey species, while primary amines were similar. Lobsters may be attracted preferentially to carrion species with higher concentrations of amines and/or higher potency of attractants per unit amine.     

Validation of Bioassays for Assessing the Contamination of Marine Environments

Various biomarkers were used to determine the exposure of fish (Arius felis and Micropogon undulatus) from Galveston Bay (GB), Texas, USA to organic contaminants. Sediment levels of polynuclear aromatic hydrocarbons (PAHs) in GB ranged from 81 to >1000 ng/g and polychlorinated biphenyls (PCBs) were <20 ng/g at all stations. No significant differences in hepatic concentrations of contaminants and ethoxyresorufin O–deethylase (EROD) activity, CYPIA mRNA levels, and DNA adducts were found in A. felis from GB. However, significant differences in biliary concentrations of naphthalene, phenanthrene, and benzo[a]pyrene metabolites were observed. Induced EROD activities and elevated levels of biliary PAH metabolites were measured in M. undulatus from the two most contaminated sites in GB. Induction toxic equivalents (I-TEQs), derived from dosing rat hepatoma H4IIE cells with hepatic extracts of A. felis, were correlated with tissue levels of ≥4–ring PAHs.     

Antifungal leaf-surface metabolites correlate with fungal abundance in sagebrush populations

A central component in understanding plant–enemy interactions is to determine whether plant enemies, such as herbivores and pathogens, mediate the evolution of plant secondary metabolites. Using 26 populations of a broadly distributed plant species, sagebrush (Artemisia tridentata), we examined whether sagebrush populations in habitats with a greater prevalence of fungi contained antifungal secondary metabolites on leaf surfaces that were more active and diverse than sagebrush populations in habitats less favorable to fungi. Because moisture and temperature play a key role in the epidemiology of most plant–pathogen interactions, we also examined the relationship between the antifungal activity of secondary metabolites and the climate of a site. We evaluated the antifungal activity of sagebrush secondary metabolites against two fungi, a wild Penicillium sp. and a laboratory yeast, Saccharomyces cerevisiae, using a filter-paper disk assay and bioautography. Comparing the 26 sagebrush populations, we found that fungal abundance was a good predictor of both the activity (r ² = 0.36 for Saccharomyces, r ² = 0.37 for Penicillium) and number (r ² = 0.34 for Saccharomyces) of antifungal secondary metabolites. This suggests that selection imposed by fungal pathogens has led to more effective antifungal secondary metabolites. We found that the antifungal activity of sagebrush secondary metabolites was negatively related to average vapor pressure deficit of the habitat (r ² = 0.60 for Saccharomyces, r ² = 0.61 for Penicillium). Differences in antifungal activity among populations were not due to the amount of secondary metabolites, but rather to qualitative differences in the composition of antifungal compounds. Although all populations in habitats with high fungal prevalence had secondary metabolites with high antifungal activity, different suites of compounds were responsible for this activity, suggesting independent outcomes of selection on plants by fungal pathogens. The location of antifungal secondary metabolites on the leaf surface is consistent with their putative defense role, and we found no evidence supporting other functions, such as protection from ultraviolet light or oxidation. That the antifungal activity of sagebrush secondary metabolites was similar for two different fungi provides support for broad antifungal defenses. The incidence and severity of fungal disease in the field (caused by Puccinia tanaceti) were similar in moist and dry habitats, possibly reflecting an equilibrium between plant defense and fungal attack, as sites with greater fungal abundance compensated with more effective secondary metabolites. The geographic correlation between fungal abundance and antifungal secondary metabolites of sagebrush, coupled with our other data showing heritable variation in these metabolites, suggests that pathogenic fungi have selected for antifungal secondary metabolites in sagebrush.     

Natural Products Isolated From Species of Halgerda Bergh, 1880 (Mollusca: Nudibranchia) and Their Ecological and Evolutionary Implications

Chemical investigations of five species of molluscan nudibranchs, Halgerda, collected from Australia and Japan were carried out. We identified a novel tryptophan derivative halgerdamine (1) along with the known compounds trigonellin (3), esmodil (4), zooanemonin (5), and C2-α-D-mannosylpyranosyl-L-tryptophan (2) from Halgerda aurantiomaculata. C2-α-D-mannosylpyranosyl-L-tryptophan (2) was previously thought to be a by-product of tryptophan metabolism exclusive to humans. The only other chordates reported to contain this compound are a number of ascidian species from New Caledonia and Australia including Atriolum robustum and Leptoclinides dubius. The occurrence of 2 in a mollusc has not been previously reported. Structure elucidation of 1 was achieved by using high-field 2D NMR spectroscopy. No secondary metabolites were detected in extracts from five of the six other species studied, whereas Halgerda gunnessi contained mixtures of acylated tetrasaccharides. The compounds isolated from Halgerda are different from those found in the close sister taxon, Asteronotus. Specifically, species of Halgerda contain no terpenes, spirodysin, or bromophenols, as does Asteronotus. Furthermore, in contrast to other members of the Nudibranchia such as Chromodoris and Phyllidia, in which compound yields are quite high, several cryptic species of Halgerda seem to lack secondary metabolites, whereas the two highly conspicuous species yield mildly cytotoxic MeOH extracts. Our findings support recent hypotheses regarding progressive evolution of opisthobranchs. In particular, opisthobranchs have evolved to exploit a wider range of food and metabolites than did their ancestors, a demonstration of physiological innovation. Some species of Halgerda may not be chemically protected either via de novo synthesis or by sequestering particularly toxic compounds.     

Phenolic Metabolites in Leaves of the Invasive Shrub, Lonicera maackii, and Their Potential Phytotoxic and Anti-Herbivore Effects

Lonicera maackii is an invasive shrub in North America for which allelopathic effects toward other plants or herbivores have been suspected. We characterized the major phenolic metabolites present in methanol extracts of L. maackii leaves. In addition, we examined the effects of methanol–water extracts of L. maackii leaves on seed germination of a target plant species and on feeding preference and growth rate of a generalist insect herbivore. A total of 13 individual major and minor compounds were detected in crude leaf extracts by high-performance liquid chromatography coupled to electronspray ionization-tandem mass spectrometry (ESI-MS/MS). Extracts were dominated by two major flavones, apigenin and luteolin, and their glucoside derivatives, apigenin-7-glucoside and luteolin-7-glucoside. Quantities of these compounds, along with chlorogenic acid, varied between two sampling points. Leaf extracts that contained these compounds were inhibitory to seed germination of Arabidopsis thaliana. In addition, treatment of artificial diet with leaf extracts deterred feeding of the generalist herbivore, Spodoptera exigua, in choice experiments but had no effect on growth rate in short-term no-choice bioassays. Purified apigenin tended to deter feeding by S. exigua and inhibited seed germination of A. thaliana. We conclude that leaves of L. maackii contain phenolic compounds, including apigenin and chlorogenic acid, capable of having biological effects on other plants and insects.     

Seasonal variation in volatile secondary compounds ofChrysothamnus nauseosus (Pallas) britt.; asteraceae ssp.hololeucus (Gray) hall. & clem. Influences herbivory

Chrysothamnus nauseosus (rubber rabbitbrush) is used by browsing animals, especially mule deer (Odocoileus hemionus), as a forage in the winter months. It is used only slightly, if at all in the summer. This dietary difference may result from changes in the secondary chemical composition of the leaves. Solvent extracts from summer and winter rabbitbrush leaves were analyzed by gas chromatography and mass spectrometry, and the volatile compounds were quantified and identified. Hexane and chloroform extracts from winter leaves exhibit a marked concentration decrease in most chemicals when compared to summer extracts. The methanol extracts revealed the presence of several chemicals in the summer leaves that were absent in winter leaves. Rubber rabbitbrush has fewer secondary volatile chemicals in the winter than in the summer. These chemical differences may influence the seasonal dietary difference observed in mule deer and other browsing animals.     

Quantitative assessment of natural toxicity in sponges: toxicity bioassay versus compound quantification

Microtox^® assay was used to assess the natural toxicity of two sponges, Dysidea avara and Ircinia variabilis. The activity of crude extracts and major metabolites were compared. Methanol extract of D. avara was more toxic than that of acetone and was as toxic as pure avarol, thus suggesting that the toxicity of the sponge was mainly due to this metabolite. We also quantified palinurin, the major metabolite of I. variabilis, in specimens from several habitats. With the same methanol extracts used for palinurin quantification, we ran the Microtox^® assay and found a positive significant regression between toxicity and concentration of this metabolite. Pure palinurin was tested at the same concentration present in the extract, and the toxicity recorded was higher than that of the methanol extract. As with avarol from D. avara, palinurin is the main secondary metabolite that confers toxicity to I. variabilis. The results confirm that the standardized Microtox^® assay is an accurate and reproducible tool for assessing the toxicity of crude extracts and pure metabolites of marine organisms. These results also suggest that methanol is more suitable than acetone for the detection of species toxicity by Microtox^® The method is faster and easier to perform than chemical quantification even when the sponge chemistry is known, and is appropriate for studies on variation in natural toxicity over a range of environmental conditions.     

(2S,12Z)-2-Acetoxy-12-heptadecene: Major Sex Pheromone Component of Pistachio Twig Borer, Kermania pistaciella

The sex pheromone of the pistachio twig borer, Kermania pistaciella (Lepidoptera: Oinophilidae), one of the most important insect pests of pistachio, Pistacia vera, in Turkey and Iran, was identified. In gas chromatographic-electroantennographic detection (GC-EAD) and GC-mass spectrometric analyses of pheromone gland extracts of female K. pistaciella from Turkey, (2S,12Z)-2-acetoxy-12-heptadecene was identified as the major candidate pheromone component. In field experiments in Turkey, lures containing synthetic (2S,12Z)-2-acetoxy-12-heptadecene attracted large numbers of male moths. Its attractiveness was significantly reduced by the presence of the R-enantiomer or of either enantiomer of the corresponding alcohol. (2S,12Z)-2-Acetoxy-12-heptadecene is the first pheromone component identified in the Oinophilidae and the first secondary acetate pheromone component identified in the Lepidoptera. An erratum to this article can be found at     

Palatability of Macroalgae that Use Different Types of Chemical Defenses

This study compared algal palatability and chemical defenses from subtropical green algae that may use different types of defense systems that deter feeding by the rock-boring sea urchin Echinometra lucunter. The potential defense systems present include (1) the terpenoid caulerpenyne and its activated products from Caulerpa spp., and (2) dimethylsulfoniopropionate (DMSP)-related defenses in Ulva spp. Secondary metabolites from these chemical groups have been shown to deter feeding by various marine herbivores, including tropical and temperate sea urchins. Live algal multiple-choice feeding assays and assays incorporating algal extracts or isolated metabolites into an artificial diet were conducted. Several green algae, including Ulva lactuca, Caulerpa prolifera, and Cladophora sp., were unpalatable. Nonpolar extracts from U. lactuca deterred feeding, whereas nonpolar extracts from C. prolifera had no effect on feeding. Polar extracts from both species stimulated feeding. Caulerpenyne deterred feeding at approximately 4% dry mass; however, dimethyl sulfide and acrylic acid had no effect at natural and elevated concentrations. E. lucunter is more tolerant than other sea urchins to DMSP-related defenses and less tolerant to caulerpenyne than many reef fish. Understanding the chemical defenses of the algae tested in this study is important because they, and related species, frequently are invasive or form blooms, and can significantly modify marine ecosystems.     

Larval Beetles Form a Defense from Recycled Host-Plant Chemicals Discharged as Fecal Wastes

Larvae of the leaf-feeding beetles Neolema sexpunctata and Lema trilinea carry feces on their backs that form shields. We used the generalist predatory ant, Formica subsericea, in a bioassay to determine whether shields were a physical barrier or functioned as a chemical defense. Fecal shields protected both species against ant attack. Larvae of both species reared on lettuce produced fecal shields that failed to deter ants. Commelina communis, N. sexpunctata's host, lacks noxious secondary compounds but is rich in phytol and fatty acids, metabolites of which become incorporated into the fecal defense. In contrast, the host plant of L. trilinea, Solanum dulcamara, contains steroidal glycoalkaloids and saponins, whose partially deglycosylated metabolites, together with fatty acids, appear in Lema feces. Both beetle species make modifications to host-derived precursors before incorporating the metabolites into shields. Synthetic chemicals identified as shield metabolites were deterrent when applied to baits. This study provides experimental evidence that herbivorous beetles form a chemical defense by the elimination of both primary and secondary host-derived compounds. The use of host-derived compounds in waste-based defenses may be a more widely employed strategy than was hitherto recognized, especially in instances where host plants lack elaborate secondary compounds.     

Chemosensory responses of cowpea weevil,Callosobruchus chinensis to an aquatic weed,water hyacinth,Eichhornia crassipes (Mart) Solms

The behavioral responses of cowpea weevil,Callosobruchus chinensis to the leaf extracts of the aquatic weed, water hyacinth,Eichhornia crassipes (Mart) Solms. was evaluated by three different methods. The petroleum ether-soluble fraction ofE. crassipes induced high weevil attraction in all methods employed. The olfactometer assay provided a reading on weevil chemosensory responses, whereas the free-choice tests measured response and preference of weevils to the untreated or extract-treated food grain of their choice over a longer period. The extracts were also presented under no choice test conditions to assay them for the presence of oviposition stimulants. In all these methods the extracts of water hyacinth evoked a quick directional response. Another interesting feature is that the extract, besides producing attraction, also caused mortality of the insects.     

Comparative Metabolism of Dietary Terpene, p-Cymene, in Generalist and Specialist Folivorous Marsupials

The urinary metabolites of the monoterpene, p-cymene, were studied in three marsupial species: a generalist herbivore, the brushtail possum (Trichosurus vulpecula), and two specialist folivores, the greater glider (Petauroides volans) and the ringtail possum (Pseudocheirus peregrinus), as well as in the laboratory rat (Rattus norvegicus). Each animal was dosed, intragastrically, with single doses of p-cymene (0.37 mmol/kg and/or 1.49 mmol/kg). Urine and feces were collected for two 24-hr periods. Quantitative analysis of urinary metabolites by extraction, gas chromatography, and mass spectrometry gave a mean recovery of 64% (range 52–74%) of the administered dose in 48 hr for the four species. No fecal metabolites were found. A species-specific pattern of metabolite excretion was evident and reflected the natural occurrence of p-cymene (and terpenes in general) in the diet. If the metabolites excreted are grouped according to the total number of oxygen atoms added (one to four), then the rat excreted metabolites encompassing all degrees of oxidation, but predominantly a monooxygenated metabolite. The brushtail possum excreted metabolites having two, three, and four oxygen atoms added. The ringtail possum and greater glider only excreted metabolites with three or four oxygen atoms. Conjugation played a significant role in the excretion of oxidized metabolites in only the brushtail possums and the rat. These findings indicate that species encountering terpenes, such as p-cymene, in their normal diet have developed efficient oxidation pathways to eliminate them. This oxidative efficiency could also reduce the necessity for subsequent conjugation of metabolites which minimizes further demands on a nutritionally limited diet.