Histological and biochemical bases of defense mechanisms in four species of Polybranchioidea ascoglossan molluscs

The multiple defensive strategies of four Mediterranean ascoglossan molluscan species, belonging to two different Polybranchioidea families, Cyerce cristallina (Trinchese, 1881) and Caliphylla mediterranea (Costa A., 1869) (Polybranchiidae), and Ercolania funerea (Costa A., 1867) and Placida dendritica (Alder and Hancock, 1843) (Stiligeridae), were studied. E. funerea, P. dendritica and C. mediterranea were collected from the lake Fusaro (Arco Felice, Naples) in 1992. C. cristallina was collected from Capo Miseno (Bay of Naples) in 1991. C. cristallina and E. funerea easily undergo autotomy of dorso-lateral appendages (cerata) followed by an extraordinarily quick (8 to 10 d) regeneration of the latter. Histological analyses showed the presence, at the basis of both normal and regenerated cerata of these species, of a muscular sphincter which facilitates the autotomic process. C. mediterranea and P. dendritica, which do not undergo autotomy, lack this anatomical feature but feed upon algae belonging to the Chaetomorpha and Bryopsis genera, and, as shown by electron microscopy studies, retain large quantities of chloroplasts which they use as camouflage amidst algae and to escape predation. E. funerea also exploits this behavioral defense mechanism. Histological investigations also revealed in the cerata of all four species several multi-cellular mucous glands responsible for the secretion of the slime typical for these molluscs. C. cristallina, E. funerea and P. dendritica secrete large amounts of slime, whose extracts displayed ichthyotoxic activity when assayed by the Gambusia affinis ichthyotoxicity assay, while extracts of C. mediterranea slime were not toxic. A chemical analysis of the slime, mantle and cerata led to the isolation of polypropionate - and -pyrones from all species except C. mediterranea. These secondary metabolites possess structures that differ only by the degree of methylation and the geometry of double bonds of the side chain and are specifically distributed in cerata and slime of C. cristallina and E. funerea. The pyrones also display different activities in the Hydra vulgaris regeneration assay and in the G. affinis ichthyotoxicity test and, depending on their structural features and tissue distribution, are likely to play a role either as defense allomones or as supportive inducers of cerata regeneration. In conclusion, combined biological observations and histological and chemical techniques generated valuable information on the defensive behavior of these four ascoglossan species which exploit various combinations of the same behavioral and/or chemical defensive strategies and thus successfully avoid predation.     

Cnidosac morphology in dendronotacean and aeolidacean nudibranch molluscs: from expulsion of nematocysts to use in defense?

Nudibranchs (Mollusca, Gastropoda) feeding on tentacles and polyps of Cnidaria thereby ingest both latent and discharged nematocysts (NCs) along with the food mass. In eolid nudibranchs (Opisthobranchia, Aeolidacea), many of the undischarged NCs are transported to terminal cnidosacs in their body appendages (cerata) and incorporated as kleptocnidae for defense. In the present report, the occurrence and fate of NCs in the digestive tracts of eolids is compared with hydrozoan-feeding dendronotacean nudibranchs (Opisthobranchia, Dendronotacea), which may show more basic stages in the evolution of cnidosacs. Tomographic reconstructions of the distal tips of cerata were composed from series of semithin light microscopic sections, utilizing 3D-surface rendering software. Doto acuta (Dendronotacea, Dotidae) does not have cnidosacs; transmission electron micrographs show that the NCs are digested in lysosomes of digestive gland cells. In contrast, species of the genus Hancockia (Dendronotacea, Hancockiidae) have several small cnidosacs in each ceras; they accumulate NCs in the digestive cells, as well as in the cnidosacs. Many of the cnidosacs were found open to the exterior with NCs in the process of expulsion. These and other structural details suggest assigning a function of expelling the NCs to the Hancockia spp. cnidosacs. It is proposed that cnidosacs similar to those of Hancockia spp. provide a clue to understanding how the defensive function of eolid cnidosacs may have evolved. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Ecological investigations on the zooplankton community of Balsfjorden,northern Norway: Lipids in the euphausiids Thysanoessa raschi and T. inermis during spring

Dry body weights, lipid levels and lipid compositions were measured in I-and II-group Thysanoessa raschi (M. Sars) and T. inermis (Krøyer) collected in April-May, 1980 in Balsfjorden, north Norway. Dry body weights were mininal in late April but had doubled by mid-May for I-group T. raschi and for I- and II-group T. inermis. II-group T. raschi had increased its dry weight by less than 50% by mid-May. Lipid accounted for approximately 10% of the dry body weight throughout the period, with free fatty acids and phospholipids being the dominant classes. An exception occurred for II-group T. inermis in mid-May, when wax esters were present in substantial amounts. Wax esters were present only in small amounts at other times and triacylglycerols were negligible. The free fatty acids in I-group T. raschi and I-group T. inermis in late April were deficient in polyunsaturates; by mid-May the free fatty acids were rich in polyunsaturates. II-group T. raschi and II-group T. inermis had free fatty acids rich in polyunsaturates throughout the period of study. The wax esters present in II-group T. inermis in mid-May consisted mainly of 16:0 and 14:0 alcohols esterified to 18:1 fatty acid. The traces of wax esters present in T. raschi did not contain significant amounts of phytol. Results are discossed with respect to the metabolic activities of the two euphausiids and their trophic positions in terms of different dietary inputs.     

Protein and amino acid composition of the tergal gland secretions ofBlatta orientalis andEurycotis floridana (Dictyoptera: Blattidae)

Summary Nymphs and adult females ofBlatta orientalis and nymphs ofEurycotis floridana produce a proteinaceous sticky secretion which accumulates on the last abdominal tergites. The proteic patterns do not differ between individuals of the same species. HPLC analyses show that all the common amino acids are found in both species, aspartic and glutamic acids representing 24 to 37% of the total amount of amino acids. InB. orientalis, glutamic acid is the more abundant amino acid whereas inE. floridana it is the aspartic acid. The secretion appears and accumulates rapidly on isolated insects. Behavioural assays revealed that these secretions have a defensive role.     

Management of nematocysts in the alimentary tract and in cnidosacs of the aeolid nudibranch gastropod<Emphasis Type="Italic"> Cratena peregrina</Emphasis>

The nudibranch gastropod Cratena peregrina (Opisthobranchia: Aeolidaceae), when feeding on polyps of the hydrozoan Eudendrium racemosum (Cnidaria: Anthoathecata), devours masses of small microbasic eurytele and holotrichous isorhiza nematocysts. Large proportions of these nematocysts were found undischarged in the alimentary tracts of the snails. Feeding experiments in this study tracked the fate of nematocysts as they passed through the alimentary canal to the digestive gland in the dorsal appendages, the cerata, to the cnidosacs, and finally in the faeces. In digestive cells, many structurally intact nematocysts were present in large phagosomes that remained unaffected even after 2 days fasting, and phagosomes containing nematocysts were found in the faeces. Thus, it is inferred that fusion of nematocyst-containing phagosomes with lysosomes and subsequent digestion of nematocysts is blocked. Masses of exposed, undigested and structurally intact nematocysts were discarded in the faeces. In the tips of the cerata, other nematocysts were phagocytosed by cnidophages and stored in the cnidosacs. After release, in contact with seawater, cnidosac nematocysts were able to discharge. When cnidophore tentacles of E. racemosum with only holotrichous isorhizas were fed to the snails, the isorhizas arrived in the cnidosacs about 2 h after feeding and mixed with existing small euryteles. Some cnidosacs also contained very large microbasic eurytele or large macrobasic eurytele nematocysts, possibly from Eudendrium ramosum and Eudendrium glomeratum, respectively. This indicates that the various types of nematocysts from food were all incorporated into the cnidosacs. Evidence for a selection process or digestion of a certain nematocyst type in the cnidophages was not obtained. It is concluded that a large proportion of the nematocysts ingested with the food are not digested, but are eliminated, structurally and functionally intact, via the alimentary canal and the tips of the cerata.     

Fish-bone oil: Percent total body lipid and carbon-14 uptake following feeding of 1-14C-palmitic acid

Bones contain the majority of body lipid in some marine fish. In the sheepshead wrasse Pimelometopon pulchrum and the sablefish Anoplopoma fimbria, the bone lipid comprised 79 to 93% and 52 to 82% of the total body lipid, respectively. The senorita Oxyjulus california, another species of wrasse, has only 14% of its body lipid in bone. To determine whether dietary lipid is deposited quickly in the bone lipid, three species of fish were fed 1-¹⁴C-palmitic acid. Radioactivity appeared in the bone lipid as soon as 12 h after feeding, with the species rich in bone lipid incorporating the highest activity relative to the flesh. Roughly 80% of the radioactivity recovered in A. fimbria bone lipid was found in triglyceride. Radioactivity was equally distributed between phospholipid and triglyceride in P. pulchrum bone lipids. The results suggest that in some marine fish the bones contain the majority of the organism's reserve energy.     

Aerial defense of the nest by workers of the stingless bee Trigona (Tetragonisca) angustula (Latreille) (Hymenoptera: Apidae)

Summary The stingless bee Trigona (Tetragonisca) angustula has a sophisticated defense strategy against flying insect predators at the entrance of its nest. Groups of worker bees hover on both sides in front of the nest entrance tube, facing a flight corridor leading to the nest. Intruders which enter this corridor are attacked by these bees from the side and from behind and are forced to the ground by biting bees clinging to their wings. T. angustula is subject to predation by Lestrimelitta limao, a cleptobiotic stingless bee which performs organized raids on other nests to rob food supplies, larval provisions and nest constructing material. The presence of citral, released by L. limao during the raids, leads to a rapid increase in the number of hovering guard bees in front of T. angustula nests. This recruitment in response to citral suggests that the defense behavior in T. angustula has evolved under the pressure of L. limao raids and that citral functions in T. angustula as an alarm kairomone.     

On the lipid biochemistry of polar copepods: compositional differences in the Antarctic calanoids Euchaeta antarctica and Euchirella rostromagna

During austral summer of 1985 different developmental stages (CIII, CIV, CV, females, males) of the Antarctic copepod Euchaeta antarctica and females of Euchirella rostromagna were collected in the southeastern Weddell Sea to determine their lipid contents and compositions. For E. antarctica the analyses revealed a strong ontogenetic accumulation of lipids towards the older copepodids with highest lipid contents in late CV stages and adults. The females of E. rostromagna had moderate lipid levels. The most striking difference between these two species concerns their lipid class compositions. E. antarctica deposited predominantly wax esters, whereas in E. rostromagna the major lipid class consisted of triacylglycerols, an unusual storage lipid in polar marine copepods. Principal fatty acids in E. antarctica were the monounsaturates 18:1(n-9) and 16:1(n-7), especially in the lipid-rich stages, while the polyunsaturated fatty acids 20:5(n-3) and 22:6(n-3), usually membrane lipids, dominated in the lipid-poor stages. The wax ester moieties in E. antarctica consisted almost entirely of 14:0 and 16:0 fatty alcohols. Major components in E. rostromagna were the fatty acids 18:1(n-9), 16:0, 20:5(n-3) and 22:6(n-3). The potential of fatty acids and alcohols as typical trophic markers is rendered largely insignificant in the two species due to catabolic processes.     

Nutritional phagostimulants function as male courtship pheromone in the German cockroach, <Emphasis Type="Italic">Blattella germanica</Emphasis>

Summary. Males of the German cockroach, Blattella germanica, secrete a pheromonal substance from the abdominal tergal glands, which elicits a feeding response in females during the sequential courtship behavior. The nuptial secretion consists predominantly of a synergistic mixture of sugars and phospholipids. Cholesterol and a series of amino acids, which are also components of the males glandular secretion, significantly enhanced the phagostimulant activity of the sugar components. The nuptial feeding behavior of the female cockroach is therefore elicited by a complex assortment of nutritive components in the male tergal secretion, including sugars, phospholipids, cholesterol, and amino acids. These results indicate that a mixture of primary metabolites, and not of specific secondary metabolites, serves as a pheromonal cue that appeals to the females gustatory sense and effectively brings her to the precopulatory position. Although the male secretion consists of nutrients, we suggest that these compounds probably do not represent a significant nutrient investment in females and their progeny but rather function as a signal in the mating sequence of B. germanica.     

Ultrastructural analysis of dissolution of shell of the bivalve Mytilus edulis by the accessory boring organ of the gastropod Urosalpinx cinerea

The shell penetrating mechanism of the muricid gastropod Urosalpinx cinerea follyensis Baker was examined with reference to solubilizing effects of the secretion of the accessory boring organ (ABO) on the ultrastructural organic and mineral components of the shell of the bivalve Mytilus edulis Linné. The fine structure of shell etched by the secretion was contrasted with normal shell and shell solubilized artificially by acids, a chelating agent, and enzymes as an aid in interpreting the pattern etched by the secretion. A synoptic series of scanning electron micrographs of representative regions of the normal shell of M. edulis was prepared to serve as a standard for ultrastructural interpretation of the patterns of dissolution. Intercrystalline conchiolin of the mosaicostracal, prismatic, myostracal, and nacreous strata was dissolved as readily as the periostracum by the ABO secretion. In the prismatic region, maximum depth of dissolution of intercrystalline organic matrix occurred when long axes of prisms were approximately perpendicular to the surface being dissolved. Microscopic solubilization of organic matrix noticeably preceded dissolution of mineral crystals, revealing subsurface prisms and lamellae similar in size and form to the distinctively shaped normal shell units. After solubilization of intercrystalline conchiolin, further dissolution by the ABO secretion revealed a variety of what appeared like internal structures in prisms and lamellae. The form of these subunits varied from that of platelets to nodules in prisms and from laths to nodules in lamellae. These intracrystalline patterns of dissolution probably resulted from preferential etching of (a) soluble intracrystalline conchiolin membranes or other internal aggregates of nacrin, (b) heterogeneously distributed trace and minor elements, or (c) from both. Carriker and Williams (1978) hypothesized that a combination of HCl, chelating agents, and enzymes in a hypertonic mucoid secretion released by the ABO dissolve shell during hole boring. The similarity of patterns of dissolution etched by the ABO secretion and those produced artificially by HCl and EDTA in the present study support the hypothesis that these chemicals, or chemicals similar to them, are constituents of the ABO secretion. Lactic and succinic acids and a chitinase-like enzyme were also suggested by Carriker and Williams as possible agents in shell dissolution. Alteration of the shell surface by experimental application of these, and other, chemical agents was not sufficiently comparable to that etched by the ABO secretion to support the suggestion. Preferential dissolution of shell matrix by the ABO secretion is functionally advantageous to boring gastropods because it increases the surface area of mineral crystals exposed to solubilization and facilitates removal of shell units from the surface of the borehole by the radula.     

Responses of ovaries and testes of Lytechinus variegatus (Echinodermata: Echinoidea) to dietary administration of estradiol, progesterone and testosterone

 In many vertebrates, environmental factors influence gamete differentiation and growth of the mature gonad through alteration of sex steroid production or action; however, it is unclear how gamete differentiation and gonadal growth are regulated in echinoids. The purpose of this study was to examine the influence of dietary administration of estradiol (E2), progesterone (P4), testosterone (T) and finasteride (F, a 5α-reductase inhibitor) on the ovaries and testes of mature Lytechinus variegatus (Lamarck) during gonadal growth. Echinoids were fed a formulated diet supplemented with steroids or steroids in combination with finasteride for 36 d. The effects of dietary administration of steroids on L. variegatus were both steroid- and sex-specific. The mean ovary index was 54% greater in individuals fed E2 than from individuals fed the control (C) diet (10.0 ± 1.1 vs 6.5 ± 0.7, respectively; P < 0.05). Individuals fed E2, P4, E2/P4, and P4 in combination with F had significantly smaller oocytes (P < 0.005) than individuals fed C. The volume fraction occupied by nutritive phagocytes in ovarian tubules from individuals fed E2/P4 or P4 were significantly larger than the volume fraction occupied by nutritive phagocytes from individuals fed C, indicating that E2 and 5α-reduced progestins may promote nutrient allocation to nutritive phagocytes. Although oocytes from the individuals fed T alone were significantly smaller than those fed T in combination with F, oocytes from individuals in both treatments were significantly larger compared to oocytes from individuals fed C. These data suggest that upon removal of 5α-reduced androgens, T is able to promote an increase in oocyte diameters. In contrast, the mean testis index was 56% greater in individuals fed P4 than in individuals fed C (8.9 ± 0.6 vs 5.7 ± 0.9, respectively; P < 0.05); the testis index did not increase in individuals fed P4/F, suggesting that 5α-reduced progestins stimulate testicular growth. Testes growth in the presence of 5α-reduced progestins was accomplished by significant increases in the volume fraction occupied by nutritive phagocytes and by a significant reduction in the volume fraction occupied by spermatogenic columns in testicular tubules. These data further indicate that 5α-reduced progestins (or 5α-reduced androgens) may inhibit spermatogenic column formation. In conclusion, E2 stimulated ovarian growth but inhibited oocyte growth, whereas T had no affect on ovarian growth but promoted oocyte growth in L. variegatus. We hypothesize that the E2 (or E2 metabolites) and/or 5α-reduced androgens in combination with T regulate oocyte growth in the echinoid L. variegatus. In addition, 5α-reduced progestins promoted nutrient accumulation in nutritive phagocytes within the ovaries and the testes. Furthermore, 5α-reduced progestins stimulated growth of the testes and inhibited spermatogenic column formation, suggesting that 5α-reduced progestins regulate nutrient accumulation into nutritive phagocytes and spermatogenic column formation in L. variegatus. The differences in estrogen effects between echinoids and asteroids may be related to differences in gonad morphology and, ultimately, the differences in cellular signalling pathways (paracrine vs endocrine). Received: 22 May 1999 / Accepted: 24 May 2000     

Growth and grazing rates of the tintinnid ciliate<Emphasis Type="Italic"> Favella taraikaensis</Emphasis> on the toxic dinoflagellate<Emphasis Type="Italic"> Alexandrium tamarense</Emphasis>

Growth and feeding activities of the tintinnid ciliate Favella taraikaensis fed the toxic dinoflagellate Alexandrium tamarense were examined in laboratory experiments. Both growth and ingestion rates of F. taraikaensis as a function of the A. tamarense concentration were fitted to a rectangular hyperbolic equation. The maximum growth and ingestion rates of F. taraikaensis were 1.0 day^–1 and 2.8 cells ind. h^–1 (carbon specific ingestion rates: 3.5 day^–1), respectively, which are both included in the range of previous data reported for Favella spp. feeding on other algae. The gross growth efficiency (GGE) of F. taraikaensis ranged from 0.26 to 0.49 (mean value 0.40) at the concentration of 10–800 cells ml^–1, which is within the range of previous data on Favella spp. Also, the growth and ingestion rates and GGE of F. taraikaensis on A. tamarense were not significantly different from the values on another non-toxic dinoflagellate (Heterocapsa triquetra) at two different prey concentrations. This indicates that the toxicity of A. tamarense probably did not influence the feeding and growth activities of F. taraikaensis at concentrations of less than ca. 800 cells ml^–1. To evaluate the grazing by F. taraikaensis on A. tamarense blooms in the field, the population dynamics of A. tamarense were simulated based on the growth and ingestion parameters of F. taraikaensis. As a result, the grazing impact by F. taraikaensis was considered to potentially regulate the development of A. tamarense blooms. If the toxicity of A. tamarense does not influence the growth and feeding activities of F. taraikaensis, the occurrence of such grazer plankton are considered to be important for predicting the course of A. tamarense bloom dynamics under natural conditions.Communicated by T. Ikeda, Hakodate     

Sensitivity of zooxanthellae and non-symbiotic microalgae to stimulation of photosynthate excretion by giant clam tissue homogenate

Stimulation of photosynthate excretion from zooxanthellae and free-living algae by tissue homogenate of several bivalves was studied. Mantle tissue homogenate of Tridacna derasa enhanced 10-to 15-fold excretion of photosynthetically fixed carbon from freshly isolated zooxanthellae within 2 h incubation. Maximum carbon excretion was 35 to 45% of the total carbon fixed. This excretion stimulating activity was detected in the homogenates of the mantle, adductor muscle, gill, and kidney. However, no excretion stimulating activity was detected in the haemolymph. The excretion stimulation activity of mantle homogenate, directed against freshly isolated zooxanthellae from T. derasa, was higher in bivalves belonging to the Tridacnidae (T. derasa, T. crocea, T. maxima, T. squamosa, Hippopus hippopus) than in the Cardiidae (Fragum fragum, F. mundum, F. unedo), non-symbiotic bivalves (Mytilus edulis, Meretrix lusoria, Ruditapes philippinarum) or gastropods (Umbonium giganteum, Turbo argyrostoma). The mantle homogenate of T. derasa enhanced photosynthate excretion by free-living algae belonging to the Dinophyceae (Prorocentrum micans, Amphidinium carterae, and Heterocapsa triquetra) but did not enhance its excretion by free-living algae belonging to the Chlorophyceae, Cyanophyceae, Rhodophyceae, Prasinophyceae, and Haptophyceae. T. derase used in this study originated from Belau (Palau). T. crocea, T. squamosa, T. maxima, H. hippopus and F. unedo were collected at Ishigaki Island in Okinawa in 1992. F. mundum and F. fragm were collected at Okinawa Island in 1992.     

Sequestration of plant secondary compounds by butterflies and moths

Summary A number of aposematic butterfly and moth species sequester toxic substances from their host plants. Some of these insects can detect the toxic compounds during food assessment. Some pipevine swallowtails use aristolochic acids among the host finding cues during oviposition and larval feeding and accumulate the toxins in the body tissues throughout all life stages. Likewise, a danaine butterfly,Idea leuconoe, which sequesters high concentrations of pyrrolizidine alkaloids in the body, lays eggs in response to the specific alkaloid components contained in the apocynad host. Insect species sharing the same poisonous host plants may differ in the degree of sequestration of toxins. Two closely ralated aposematic geometrid moth species,Arichanna gaschkevitchii andA. melanaria, sequester a series of highly toxic diterpenoids (grayanotoxins) in different degrees, while a cryptic geometrid species,Biston robstus, does not sequester the toxins, illustrating the diversity in adaptation mechanisms even within the same subfamily. By contrast, a number of lepidopteran species store the same compounds though feeding upon taxonomically diverse plant species. A bitter cyanoglycoside, sarmentosin, was characterised from several moth species in the Geometridae, Zygaenidae and Yponomeutidae, and from the apollo butterflies,Parnassius spp. (Papilionidae), although each species feeds on different groups of plants.Interspecific similarities and differences in life history and ecology are discussed in relation to variable characteristics of sequestration of plant compounds among these lepidopteran insects.     

Defensive and chemical characterization of the froth produced by the cercopid Aphrophora cribrata

Nymphs of the cercopid Aphrophora cribrata cover themselves with a frothy exudate while ingesting sap from their preferred host plant, the eastern white pine, Pinus strobus. Chemical analyses of froth collected from A. cribrata nymphs revealed an array of metabolites belonging to five chemical classes, including fatty acid-derived alcohols, γ-lactones and a single 1-monoacylglycerol, as well as the polyol pinitol and the polyhydroxyalkanoate, poly-3-hydroxybutyrate. Bioassays showed the natural A. cribata froth, as well as a synthetic mixture comprised of representative compound classes identified therein, to be repellent to ants but largely devoid of topical irritancy in tests with cockroaches.     

Resistance of the generalist moth Trichoplusia ni (Noctuidae) to a novel chemical defense in the invasive plant Conium maculatum

Summary. Conium maculatum is an apiaceous species native to Eurasia that is highly toxic to vertebrates due to the presence of piperidine alkaloids, including coniine and γ-coniceine. More than 200 years after invading the United States this species remains mostly free from generalist insect herbivores. The presence of novel chemical defenses in the introduced range could provide invasive species with a competitive advantage relative to native plants. The cabbage looper (Trichoplusia ni) is a generalist lepidopteran found throughout the US that occasionally feeds on C. maculatum. We evaluated the toxicity of piperidine alkaloids to T. ni and determined putative resistance mechanisms, both behavioral and physiological, that allows this insect to develop successfully on C. maculatum foliage. T. ni larvae raised on diets enriched with coniine and γ-coniceine showed a decrease in consumption and longer development time, but no effects on growth were found at any alkaloid concentration. In a diet choice experiment T. ni larvae showed no avoidance of alkaloid-enriched diets, suggesting that the deterrence produced by alkaloids was related to a post-ingestive metabolic response. The ability of T. ni to consume diets high in alkaloid content could be due to at least three different mechanisms: 1) a decreased consumption rate, 2) efficient excretion of at least 1/3 of ingested alkaloids unmetabolized in frass, and 3) partial detoxification of alkaloids by cytochrome P450 s, as shown by the decreased larval growth in the presence of piperonyl butoxide, a P450 inhibitor. Even though T. ni tolerates C. maculatum alkaloids, the use of this species as a host plant could be ecologically disadvantageous due to prolonged larval growth and thus increased exposure to predators. Novel plant secondary compounds do not guarantee increased resistance to generalist herbivores.     

Introducing a new metal accumulator plant and the evaluation of its ability in removing heavy metals

A field study was conducted in a dried waste pool of a lead (Pb) mine in Arak (Iran) to find the accumulator plant(s) and to evaluate the amount of metal bioaccumulation in the root and shoot portion of the naturally growing vegetation. Concentrations of heavy metals were determined both in the soil and the plants that were grown in the dried waste pool. The concentrations of total Cu, Zn, Pb, and Ni in the waste pool were found to be higher than the natural soil and the toxic levels. The results showed that six dominant vegetations, namely, Centaurea virgata, Eleagnum angustifolia, Euphorbia macroclada, Gundelia tournefortii, Reseda lutea, and Scariola orientalis accumulated heavy metals. Based on the results, it was concluded that E. macroclada belonging to Euphorbiaceae is the best Pb accumulator and also a good accumulator for Zn, Cu, and Ni. The bioaccumulation ability of E. macroclada was evaluated in experimental pots. The study showed that the amount of heavy metals in polluted soils decreased several times during two years of phytoremediation. The accumulation of metal in the root, leaves, and shoot portions of E. macroclada varied significantly, but all the concentrations were within the toxic limits. Based on the obtained data, E. macroclada is an effective accumulator plant for soil detoxification and phytoremediation in critical conditions.     

Wavelet characterization of eucalypt flowering and the influence of climate

More robust statistical methods are required for examining phenological time series as they are often noisy and non-stationary. Wavelet analytic methods easily handle such data. A maximal overlap discrete wavelet transform (MODWT) analysis of flowering records (1940–1970) of Eucalyptus tricarpa, E. leucoxylon, E. microcarpa and E. polyanthemos identified four subcomponents in each flowering series: characterized as a non-flowering phase, duration, annual and intensity cycles. A decreasing overall trend in flowering was identified by the MODWT smoothed series. Wavelet correlation found the same contemporaneous effects of climate on flowering for Eucalyptus tricarpa and E. leucoxylon, and for E. microcarpa and E. polyanthemos. Wavelet cross-correlation analysis identified the cyclical influence of temperature and rainfall on peak flowering intensity (P < 0.0001). For each species there are 6 months of the annual cycle in which any given climate variable positively influences flowering intensity and 6 months of negative influence. For all species, rainfall exerts a negative influence when temperature is positive.     

Aristolochic acid content of South-East Asian troidine swallowtails (Lepidoptera: Papilionidae) and of Aristolochia plant species (Aristolochiaceae)

Summary. Analysis of South-East Asian troidine swallowtails revealed high variability in the content of aristolochic acids among individuals. The presence or absence of these compounds depends on the Aristolochia species available as food plant for the larvae. Only one plant species (Aristolochia philippinensis) contained a high concentration of aristolochic acids, while other species from various localities contained none or only marginal amounts. Whether aristolochic acids have a distinct function in chemical defense of these swallowtails is still an open question. Received 11 December 2000; accepted 4 August 2001.     

Neutral lipid compositions of antarctic fish tissues may reflect use of fatty acyl substrates by catabolic systems

Antarctic fishes contain large quantities of lipid in adipose tissue, blood and oxidative muscle. In this study, the fatty acyl compositions of free fatty acids (FFA) and triacylglycerol (TAG) in serum, adipose tissue and oxidative muscle were determined in two species of Antarctic fishes, Trematomus newnesi Boulenger, 1902, and Notothenia gibberifrons Lönnberg, 1905, collected off the Antarctic Peninsula in February and March 1987. Total lipid contents of serum and oxidative muscle in each species also were measured. T. newnesi, a mesopelagic species, has significantly more lipid in the serum than N. gibberifrons, a demersal species (17.6±2.4 and 10.4±1.0 mg lipid g^-1 serum, respectively). The oxidative muscle of T. newnesi also contains more lipid than does that of N. gibberifrons (12.5 and 9.3% by dry weight, respectively). Fatty acids comprising greater than 5% of one or more of the lipid pools assayed in both species include 14:0, 16:0, 16:1, 16:2, 18:0, 18:1, 20:5 and 22:6. The percentages of 16:1 were lower (p<0.05) in oxidative muscle FFA than in all other assayed lipid pools in both species, suggesting a catabolic preference for some monoenes in oxidative muscle of these fishes. The percentage of 14:0 was two-to three-fold higher in adipose tissue TAG than in all other lipid pools in both species. Differences in fatty acid compositions between lipid pools indicate than proteins involved in the mobilization, transport or catabolism of fatty acids exhibit differential recognition of fatty acids in T. newnesi and N. gibberifrons.