The chelation of metal ions by the acylpolyamine toxins from the web-spider Nephilengys cruentata: effects in the intoxication/detoxification of preys

Summary. Orb-web-spiders present a series of different strategies for prey capture, involving the use of different types of silk for web building, the use of adhesive traps in the webs, the secretion of toxic compounds to the spider’s preys in the adhesive coating of the capture web and the biosynthesis of a wide range of structurally related acylpolyamine toxins in their venoms. The polyamine toxins usually block neuromuscular junctions and/or the central nervous system (CNS) of Arthropods, targeting specially the ionotropic glutamate receptors; this way these toxins are used are as chemical weapons to kill / paralyze the spider’s prey. Polyamine toxins contain many azamethylene groups involved with the chelation of metal ions, which in turn can interact with the glutamate receptors, affecting the toxicity of these toxins. It was demonstrated that the chelation of Ni⁺², Fe⁺², Pb⁺², Ca⁺² and Mg⁺² ions by the desalted crude venom of Nephilengys cruentata and by the synthetic toxin JSTX-3, did not cause any significant change in the toxicity of the acylpolyamine toxins to the model-prey insect (honeybees). However, it was also reported that the chelation of Zn⁺² ions by the acylpolyamines potentiated the lethal / paralytic action of these toxins to the honeybees, while the chelation of Cu⁺² ions caused the inverse effect. Atomic absorption spectrometry and Plasma-ICP analysis both of N. cruentata venom and honeybee’s hemolymph revealed that the spider’s venom concentrates Zn⁺² ions, while the honeybee’s hemolymph concentrates Cu⁺² ions. These results are suggesting that the natural accumulation of Zn⁺² ions in N. cruentata venom favors the prey catching and/or its maintenance in the web, while the natural accumulation of Cu⁺² ions in prey’s hemolymph minimizes the efficiency of the acylpolyamine toxins as killing/paralyzing tool.     

Inhibitory effect of arachidonic acid on venom acetylcholinesterase

The venom of krait (Bungarus sindanus), an Elapidae snake, contains high level of acetylcholinesterase (AChE) activity. The effect of arachidonic acid on krait (Bungarus sindanus) venom AChE (acetylcholine hydrolase, EC 3.1.1.7) was studied using kinetic methods. This study evaluated for the first time whether arachidonic acid acts as potential AChE inhibitor of krait venom. Kinetic studies, using Line-Weaver Burk and Dixon plots, indicated that the inhibition produced by arachidonic acid was non-competitive, i.e., both k_m and V _max fell with increase in the concentration of arachidonic acid. The concentration of arachidonic acid required to inhibit 50% of enzymatic activity was found to be 1.9?µM while the inhibitory constant (K_i ) was 1.4?µM. The present results suggest that essential fatty acid (arachidonic acid) may also be used as an inhibitor of snake venom AChE.     

Paralytic shellfish toxins sequestered by bivalves as a defense against siphon-nipping fish

Saxidomus giganteus (butter clams), are known to sequester diet-derived paralytic shellfish toxins (PST), highly potent neurotoxins, in their siphons. Captive staghorn sculpins (Leptocotus armatus), a marine fish species known to crop bivalve siphons, developed a significant aversion to siphons from toxic but not non-toxicS. giganteus following a single conditioning feeding of toxic siphon tissues. Control fish showed no aversive response to siphons from non-toxicS. giganteus during 11 feeding sessions over 56 d. Aversive and non-aversive behavior varied with the toxicity of the siphons, but not with the geographic origin of the clams. Both experimental and control fish ate freely and showed no aversion to siphons from toxic littleneck clams (Protothaca staminea). Littleneck clams, unlikeS. giganteus, retain PST in their visceral mass but not in their siphons. Both toxic and non-toxicS. giganteus extended their siphons significantly more often and higher above the sediment surface during dark hours, but toxicS. giganteus extended their siphons higher than non-toxic individuals. These results support the hypothesis that siphon-nipping by fish may have selected for the retention of PST in butter clam siphons as a chemical defense.     

Inter-specific variation in anti-predator behavior in sympatric species of kangaroo rat

Kangaroo rats, Dipodomys, occupy desert habitats with little cover and thus are under high predation risk from diverse predators. The behavior used to assess predation risk or to escape capture is unknown. We therefore compared anti-predator behavior of two sympatric species of kangaroo rat of different sizes, D. merriami and D. spectabilis. We first examined whether kangaroo rats use olfaction as a first line of defense against snake predation and tested the rats for their responses to scent extracted from two species of snake that live sympatrically with the kangaroo rats, the Mojave rattlesnake (Crotalus scutulatus) and the gopher snake (Pitophis melanoleucus). We also tested for species differences in anti-predator behavior through 15-min interactions between the kangaroo rats and free-moving gopher snakes. We found that D. spectabilis actively approached the scent of both rattlesnakes and gopher snakes more than controls of vegetable oil and evaporated solvent (Fig. 1). In contrast, D. merriami did not differentiate snake odors from controls in the experimental arena, but they sniffed the sand where a free-moving snake had passed more than D. spectabilis. Both species successfully avoided predation in encounters with live snakes. Although total numbers of approaches and withdrawals were similar (Fig. 2), D. spectabilis spent significantly more time within striking distance of the snake than D. merriami. D. spectabilis approached the head of the snake in 93% of its approaches and often engaged in nose to snout contact with the snake. If the snake struck, D. spectabilis jumped directly backward to avoid a strike and footdrummed at a safe distance. In contrast, D. merriami oriented to the snake more than D. spectabilis, but approached the head in only 41% of the approaches and rarely engaged in nose-to-snout contact. The snakes struck, hissed and decreased predatory approaches with D. spectabilis but not with D. merriami (Fig. 3). These results show that kangaroo rats can behaviorally influence the risk of being preyed on by snakes. The two species differ, however, in how they react to snakes. The larger D. spectabilis confronts snakes while the smaller D. merriami monitors snakes from a safe distance and avoids them.     

Comparative study on paralytic shellfish toxin profiles of the dinoflagellate Gymnodinium catenatum from three different countries

Paralytic shellfish toxin profiles of the dinoflagellate Gymnodinium catenatum Graham were investigated as a possible biochemical marker to distinguish different geographic populations of this species. Isolates obtained between 1986 and 1988 from Japan, Tasmania (Australia) and Galicia (Spain) were cultured under similar conditions and the toxins produced were analyzed using HPLC. Variations in temperature, salinity, and nitrate and phosphate levels in the culture medium had no significant effect on the toxin profile, suggesting that toxins can be used as a stable biochemical marker for this dinoflagellate. All the isolates produced mainly toxins of the N-sulfocarbamoyl group (C1–C4, gonyautoxins 5 and 6) but their relative abundance differed according to their geographic origin. Furthermore, only the Australian population produced the newly found 13-deoxydecarbamoyl toxins, and these could readily be used to distinguish the Australian populations from those of the other two countries.     

Assessing the Potential Impact of Cane Toads on Australian Snakes

Abstract:   Cane toads ( Bufo marinus ) are large, highly toxic anurans that were introduced into Australia in 1937. Anecdotal reports suggest that the invasion of toads into an area is followed by dramatic declines in the abundance of terrestrial native frog-eating predators, but quantitative studies have been restricted to nonpredator taxa or aquatic predators and have generally reported minimal impacts. Will toads substantially affect Australian snakes? Based on geographic distributions and dietary composition, we identified 49 snake taxa as potentially at risk from toads. The impact of these feral prey also depends on the snakes' ability to survive after ingesting toad toxins. Based on decrements in locomotor (swimming) performance after ingesting toxin, we estimate the LD₅₀ of toad toxins for 10 of the at-risk snake species. Most species exhibited a similar low ability to tolerate toad toxins. Based on head widths relative to sizes of toads, we calculate that 7 of the 10 taxa could easily ingest a fatal dose of toxin in a single meal. The exceptions were two colubrid taxa (keelbacks [  Tropidonophis mairii ] and slatey-grey snakes [  Stegonotus cucullatus ]) with much higher resistance to toad toxins (up to 85-fold) and one elapid (swamp snakes [  Hemiaspis signata ]) with low resistance but a small relative head size and thus low maximum prey size. Overall, our analysis suggests that cane toads threaten populations of approximately 30% of terrestrial Australian snake species.     

Distribution patterns of sea urchins and barrens in shallow Mediterranean rocky reefs impacted by the illegal fishery of the rock-boring mollusc<Emphasis Type="Italic"> Lithophaga lithophaga</Emphasis>

Shallow rocky habitats in SW Apulia (SE Italy, Mediterranean Sea) were surveyed in late spring 2002 to assess distribution patterns of sea urchins (Paracentrotus lividus and Arbacia lixula) and barren habitats (coralline barrens and bare substrates) in rocky reefs impacted by the destructive fishery of the rock-boring date-mussel Lithophaga lithophaga. Sea urchin density, test size-structure and biomass, and the percent cover of barrens were evaluated at four locations (5–6 km apart from each other), two heavily impacted by the date-mussel fishery and two controls. Sea urchin density and barren habitat cover were assessed at two and three sites (100–300 m apart), respectively, within each location. Sea urchin biomass was evaluated only at the scale of locations. Average density of P. lividus did not significantly change between impacted locations and controls, whereas A. lixula showed a greater density at the impacted locations. Distribution patterns of A. lixula, in addition, differed at the spatial scale of a few metres between impacted locations and controls, being generally more aggregated at the controls. The size-frequency distribution (test diameter) of P. lividus showed a mode at 3–4 cm at the impacted locations compared to a mode at 2–3 cm in the controls. The size-frequency of A. lixula was bimodal at the damaged locations (with modes at 1–2 and 4–5 cm, respectively) and unimodal (with the mode at 4–5 cm) at the controls. Average biomass of both sea urchins (P. lividus and A. lixula) was two- to fourfold greater at the impacted locations (~600 g wet wt m^–2) than at the controls (150–250 g wet wt m^–2). Barren habitats had a far greater average cover (mainly of macroalgae) at the impacted locations (from 79% to 96%) than at control locations (from 7% to 21%). These results show that the date-mussel fishery may have the potential to affect distribution patterns of sea urchins and to greatly enhance the percent cover of barren grounds in shallow Mediterranean rocky reefs.Communicated by R. Cattaneo-Vietti, Genova     

Allozyme and mitochondrial DNA evidence of population subdivision in the purple sea urchin Strongylocentrotus purpuratus

Despite high potential for dispersal, the purple sea urchin Strongylocentrotus purpuratus was found to have significant genetic subdivision among locations. Ten geographic locations along the coast of California and Baja California were sampled between 1994 and 1995. Samples from some locations included both adult and recruit urchins. Allozyme analyses revealed a genetic mosaic, where differentiation over short geographic distances could exceed differentiation over much larger distances. Significant allozyme differentiation was found among subpopulations of adults (standardized variance, F _ST =0.033), among subpopulations of recruits (F _ST =0.037), and between adults and recruits from the same location. DNA-sequence data for the mitochondrial cytochrome oxidase I gene also showed significant heterogeneity among locations, with a mild break in haplotype frequencies observed 300 km south of Point Conception. California. Repeated sampling over time is necessary to determine whether these patterns of differentiation are stable and to begin to understand what forces produce them.     

Footdrumming and other anti-predator responses in the bannertail kangaroo rat (Dipodomys spectabilis)

Summary Anti-predator behaviors of the bannertail kangaroo rat (Dipodomys spectabilis) toward snake predators were investigated. We induced responses by presenting a live (tethered) snake and an inflatable snake decoy to rats of known ages and sex in their territories. Comparisons of behaviors during spontaneous activity (baseline), naturally occurring snake-rat interactions, and experimental procedures further clarified anti-predator behaviors. The tethered snake immediately induced high rates of footdrumming (the repeated striking of the hind feet on the ground) in all rats (n=24) (Table 2), and individuals of both sexes and all ages footdrummed significantly more at the tethered snake than at the decoy. Other antipredator behaviors included alert postures, jumpback, kicking sand and avoidance. Juvenile rats exhibited more avoidance behavior and alert postures than adults and footdrummed at the decoy only if they experienced the snake first. We conclude that footdrumming functions as an individual alarm signal against predation by snakes in D. spectabilis. The drumming probably informs a snake that it has been detected and may cause it to leave. Because rats did not footdrum in response to aerial predators (owls), we suggest that kangaroo rats use different anti-predator strategies that depend on the kind of threat and the type of predator.     

Introduced versus native subspecies of Codium fragile: how distinctive is the invasive subspecies tomentosoides?

After its introduction, the green alga Codium fragile (Sur.) Hariot ssp. tomentosoides (van Goor) Silva has spread widely on several temperate-zone, rocky shores where non-weedy conspecific subspecies occur (N.E. Atlantic, N.E. Pacific, S. Pacific). To determine how phenologically and morphologically distinctive the invasive alga was relative to native subspecies, I compared marine intertidal populations of C. fragile ssp. tomentosoides and the native C. fragile ssp. novaezelandiae (J. Ag.) Silva (hereafter referred to as ssp. tomentosoides and ssp. novae-zelandiae respectively on New Zealand shores in 1992, 1993 and 1995. On the North Island, the invasive ssp. tomentosoides is sparsely distributed on low intertidal benches on wave-protected shores in the Hauraki Gulf (east coast) in spring and summer, and thalli die back to the perennial holdfast in autumn. In contrast, the native ssp. novaezelandiae forms dense beds within the low intertidal mussel zone on wave-swept shores of Maori Bay (west coast), and fronds are perennial. Whereas ssp. tomentosoides has only a few fronds arising from the spongy basal hold-fast, ssp. novae-zelandiae thalli are composed of many fronds. The ssp. tomentosoides from the Hauraki Gulf is significantly more branched than comparably sized native conspecifics from Maori Bay. These phenological and morphological differences were used to predict the subspecific identity of C. fragile from three other locations on the North Island, two locations on the South Island, and four locations on S.E. Australian shores; microscopic examination of utricles was used to check the predictions. Seasonality and number of fronds per thallus are the most reliable characters for field identification of native vs invasive subspecies: perennial intertidal thalli with large numbers of fronds are indicative of native subspecies for different geographic regions.     

Effects of nutrient limitation on toxin production and composition in the marine dinoflagellate Protogonyaulax tamarensis

Toxin production was measured by high pressure liquid chromatography (HPLC) when the marine dinoflagellate Protogonyaulax tamarensis (NEPCC 255) was grown under nitrogen or phosphorus limitation. The major toxins found in P. tamarensis (255) consisted of (N21-SO ₃ ^- )STX (11%), (N21-SO ₃ ^- )NeoSTX (44%), and [(N21-SO ₃ ^- )GTX₂ plus (N21-SO ₃ ^- )GTX₃] (20%). Total toxin content on a per cell basis was high for cultures in log phase (30 to 40 fmol cell^-1) and then decreased to ca 20 fmol cell^-1 as the cultures entered stationary phase. There was a gradual decrease in the toxin content per cell during nitrogen-limited stationary phase to ca 3 fmol cell^-1 or less. Phosphorus-limited cultures showed a markedly different response than nitrogen-limited cultures. Toxin content in P-limited cells dramatically increased at the start of stationary phase, reaching levels 3 to 4 times that observed in control and nitrogen-limited cultures. These results cannot be explained by changes in the average cell volume. Eventhough dramatic effects on the total toxin concentration were observed in response to nutrient limitation (N or P), the toxin composition (on a percent basis) remained constant. This suggests that the individual toxin composition of a given isolate is a fixed genetic trait and not a transient response to changing environmental factors.     

Paralytic shellfish poison in the “hiogi” scallop Chlamys nobilis

Attempts were made to analyze the toxin composition of the toxic hiogi scallop Chlamys nobilis. The toxins were partially purified from the digestive glands by column chromatography using Bio-Gel P-2 and Bio-Rex 70 (H⁺ form), resulting in separation into protogonyautoxin (PX), gonyautoxin (GTX) and saxitoxin (STX) fractions. Their total potencies were scored to be <100 mouse units (MU), 3200 MU and 3700 MU, respectively. A 5-min hydrolysis with 0.1 N HCl enhanced the potencies of PX and GTX fractions to 450 MU and 12000 MU respectively, whereas no enhancement occurred in the STX fraction at all. Electrophoretic, thin-layer chromatographic and high performance liquid chromatographic analyses demonstrated that the PX fraction consisted mainly of GTX₈ and its epimer, the GTX fraction of GTX₅, GTX₆, along with two unidentified toxins, and the STX fraction exclusively of two unidentified toxins. This rather unique composition suggested a complex metabolism of PSP in this species.     

Metabolism and body composition of a copepod (<Emphasis Type="Italic">Neocalanus cristatus</Emphasis>: Crustacea) from the bathypelagic zone of the Oyashio region,western subarctic Pacific

Metabolism [respiratory oxygen consumption, electron-transfer-system (ETS) activity] and body composition [water, ash, carbon (C), nitrogen (N), carbon/nitrogen (C/N) ratio] of stage C5/C6 Neocalanus cristatus from 1000 to 2000 m depth of the Oyashio region, western subarctic Pacific, were determined during the period of July 2000 through June 2003. Compared with the C5 specimens from shallow depths (<250 m), those from 1000 to 2000 m were characterized by quiescent behavior, reduced respiration rates (30% of the rates at active feeding), very low water content (61–70% of wet weight), but high C content (56–64% of dry weight) and C/N ratios (7.2–10.6, by weight). Artifacts due to the recovery of live specimens from the bathypelagic zone appeared to be unlikely in this study, as judged by the consistent results between re-compression (100 atm) and non-compression (1 atm) respiration experiments, and between ETS activities and respiration rates directly measured. In addition, the respiration rates of C6 males and females of N. cristatus from the same 1000–2000 m depth were two to three times higher than the rates of C5 individuals, but were similar to the rates of a bathypelagic copepod, Paraeuchaeta rubra. Combining these results with literature data, C budgets of: (1) diapausing C5 specimens, weighing 6–10 mg dry weight; (2) molt to C6 females; and (3) the complete the life span were established, taking into account assorted losses in respiration during diapause at stages C5 and C6, molt production and egg production. Respiratory C losses by C5 and C6 specimens were estimated on the basis of body N as adjusted metabolic rates [AMR; µl O₂ (mg body N)^–0.843 h^–1], then N budgets were also computed subtracting N lost in the form of cast molts and eggs from the initial stock. Calculations revealed that allocation of the C stock was greatest to egg production (34–57%), followed by respiration (27%) and cast molts (3%), leaving residual C of 13–36% in spent C6 females. The present results for N. cristatus from the North Pacific are compared with those of Calanus spp. in the North Atlantic.Communicated by O. Kinne, Oldendorf/Luhe     

Toxicity of Brachuryan crabs in India

Crabs are the most abundant macrofauna in the coastal environment, and may belong to many different species and even families. Some of these crabs contain specialized protein substances that act as toxins. Toxins, especially proteins that are produced by living cells or organisms, are poisonous substances. Toxins can be divided according to their pharmacological effects (neurotoxins, myotoxins, hepatotoxins, nephrotoxins, hematological toxins, and locally acting toxins) and their sources, such as snakes, scorpions, mussels, spiders, marine creatures, or sea anemones. Crab toxins are heat resistant and are found in the crabs’ viscera. Hence, they cannot be destroyed by cooking, and there is also no known treatment available for these toxins. The only alternative is to educate fishermen, holiday makers, and seafood consumers to recognize these poisonous species and avoid them. Several toxins identified from crabs are neurotoxins, related to tetradotoxin (TTX), which is found in pufferfish. Other toxins include saxitoxin (STX), neurosaxitoxin (NeoSTX), and some unidentified toxins. Some poisonous crabs possess a mixture of these toxins. This article describes the toxins and Brachuryan crabs producing toxicity.     

Field depuration and biotransformation of paralytic shellfish toxins in scallop<Emphasis Type="Italic"> Chlamys nobilis</Emphasis> and green-lipped mussel<Emphasis Type="Italic"> Perna viridis</Emphasis>

Under laboratory conditions, the scallop Chlamys nobilis and the mussel Perna viridis were exposed to N-sulfocarbamoyl toxins (C2 toxin), a paralytic shellfish toxin (PST), by feeding a local toxic strain of the dinoflagellate Alexandrium tamarense (ATDP) that produced C2 toxin exclusively. The bivalves were subsequently depurated in the field, and their depuration kinetics, biotransformation and toxin distribution were quantified. Depuration was characterized by a rapid loss within the first day, followed by a secondary slower loss of toxins. In the fast depuration phase, scallops detoxified PSTs more quickly than the mussels (depuration rate constants for scallops and mussels were 1.16 day^–1 and 0.87 day^–1, respectively). In contrast, the mussels detoxified PSTs more quickly than the scallops in the slow depuration phase, and the calculated depuration rate constants (mean+SE) from day 2 to day 13 were 0.063+0.009 day^–1 and 0.040+0.019 day^–1 for mussels and scallops, respectively. The differences in the appearances of gonyautoxins, GTX2 and GTX3, and their decarbamoyl derivatives, dcGTX2, dcGTX3 and GTX5, which are all derivatives of C2 toxin, indicated active and species-specific biotransformation of the algal toxins in the two bivalves. In both species of bivalves, the non-viscera tissue contained fewer toxins and lower concentrations than the viscera-containing tissue compartment. In scallops, very little toxin was distributed in the adductor muscle. In mussels, most of the PSTs were found in the digestive gland with significant transport of toxins into the digestive gland from other tissues during the course of depuration. The toxin profiles of scallops and mussels differed from each other and from that of the toxic algae fed. A significant fraction of GTX5 was detected in the mussels but not in the scallops. Our study demonstrates a species specificity in the depuration kinetics, biotransformation and tissue distribution of PSTs among different bivalves.Communicated by T. Ikeda, Hakodate     

Do female roe deer in good condition produce more sons than daughters

In polygynous roe deer Capreolus capreolus, males are only slightly heavier than females and the overall sex ratio at birth is close to unity. We studied offspring sex ratio and litter size (range 1–4, n = 74) of culled females, in utero, which provided an opportunity to examine responses of sex ratio to maternal condition. Male embryos were heavier than their sisters, and male fawns (9 months old) heavier than female fawns, suggesting a higher growth rate in males. There was no evidence for differential mortality between the sexes from birth to 9 months old. Heavier adult females produced larger embryos than lighter, or primiparous females. The overall sex ratio of embryos did not differ from unity, but adult does had more male embryos (55%) than primiparous does (32%), and the proportion of male embryos in a litter increased with the mother's body mass. Litter size also tended to increase with maternal age and body mass. We argue that this pattern reflects adaptive variation in offspring sex ratio.     

Toxicity of the venom in three neotropical Crematogaster ants (Formicidae: Myrmicinae)

Bioassays were performed to investigate and compare the toxic properties of the contact venoms of three Neotropical Crematogaster species (C. sp. prox. abstinens, C. distans and C. brevispinosa rochai), whose main venom compounds are chemically different (long chain derivatives linked to an aldehyde or a primary acetate, and furanocembrenoid diterpenes, respectively). Different quantities of venom were topically applied on the bodies of three target insect species (workers of the termite Nasutitermes ephratae, workers of the ant Solenopsis sp., “media” and “major” workers of the ant Cephalotes pusillus). The toxicity of the venom greatly varied according to the Crematogaster species, the venom of C. b. rochai showing the highest toxic activity towards all target species. The sensitivity of the target species also greatly varied, the workers of N. ephratae showing a great sensitivity to the venoms, while the heavily armoured workers of the ant C. pusillus, particularly “major” ones, were resistant. Cuticle thickness was found to be a key factor in the resistance to the toxic effects of Crematogaster venom. Sensitivity of the target workers was also directly related to the amount of venom applied to their cuticle. Workers of C. distans were resistant to topical application of their own venom, while C. sp. prox. abstinens and C. b. rochai were not. In all cases, the venoms were slow acting, as several hours to one or even several days were needed to obtain a significant mortality. The main biological activities of the Crematogaster venom could be related to more immediate repellent properties, as recently shown with the European species C. scutellaris.     

Sperm polymorphism and genetic divergence in the mussel Perumytilus purpuratus

For the first time in a marine bivalve species, geographic intraspecific variation in sperm morphology in the Chilean mussel, Perumytilus purpuratus, is reported. Samples were obtained in December 2010 from the rocky intertidal at three locations: Antofagasta (23o44′01.08′′S, 70o26′26.11′′W), Valparaíso (32o57′23.07′′S, 71o33′01.86′′W), and Valdivia (39°50′44.26′′S, 73°25′51.99′′W). Scanning electron microscopy was used to describe sperm traits, and genetic data were obtained by sequencing a 16S mtDNA 427-bp fragment. The results showed geographic intraspecific variation with two statistically significant patterns of genetically differentiated sperm morphologies. One genetic clade was found for Antofagasta and Valparaíso, displaying a unique sperm morphotype with a short acrosome accounting for 26 % of the head length, and the other from Valdivia, with a larger acrosome occupying 47 % of the head length. These results will lead to new research to distinguish among alternative hypotheses: the presence of cryptic species, incipient speciation, or a polymorphism in sperm traits of P. purpuratus.     

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.     

Population genetics of the blue crabCallinectes sapidus: modest population structuring in a background of high gene flow

To determine the genetic population structure of blue crabs (Callinectes sapidus Rathbun), electrophoretic allozyme analysis was performed on 750 individuals collected from 16 nearshore locations ranging from New York to Texas, USA. Twenty enzymes and non-enzymatic proteins coded by 31 presumptive loci were examined. Twenty-two loci were either monomorphic or polymorphic at less than theP ₉₅ level; alleles for these polymorphic loci were geographically dispersed. Allele frequencies for three of the remaining polymorphic loci were homogeneous over all populations, as were levels of polymorphism and heterozygosity. Phenograms generated by the UPGMA (unweighted pair-group method using arithmetic averages) and distance Wagner methods exhibited no geographic pattern in the clustering of populations. Estimates ofN _em (effective number of migrants per generation between populations) indicated substantial gene flow, with aalues sufficiently high to infer panmixia between all blue crab populations from New York to Texas. However, despite this high level of gene flow, two striking patterns of geographic differentiation occurred: genetic patchiness and clinal variation. Allele frequencies atEST-2, GP-1, IDHP-2, DPEP-1, DPEP-2, andTPEP exhibited genetic patchiness on local and range-wide geographic scales, and allele frequencies atEST-2 varied temporally. Genetic patchiness in blue crabs is likely to be the result of the pre-settlement formation and subsequent settlement of genetically heterogeneous patches of larvae; allele frequencies of those larval patches may then be further modified through ontogeny by localized selection. In the Atlantic Ocean, a regional latitudinal cline ofEST-2 allele frequencies was superimposed on the range-wide genetic patchiness exhibited by that locus. This pattern against a background of high gene flow is highly likely to be maintained by selection. In estuaries along the Atlantic Ocean coast, a combination of low adult long-distance migration and a high retention rate of locally spawned larvae could serve to segregate populations and allow for the development of the geographic cline inEST-2. The Gulf of Mexico showed no apparent cline, perhaps due to long-distance migration of females in some regions of the Gulf, or to masking by genetic patchiness. These results emphasize the importance of both ecological and evolutionary time scales and structuring mechanisms in determining genetic population structure.