Metabolic transformations of acquired lucibufagins by firefly “femmes fatales”

Summary. Female Photuris fireflies sequester defensive steroidal pyrones (lucibufagins) from male fireflies of the genus Photinus. Lucibufagin analyses of Photuris females and Photinus ignitus males show that the lucibufagin mixtures of predator and prey differ in their composition. Analyses of whole body extracts showed that P. ignitus males contain a mixture of eight non-glycosylated lucibufagins, composed mostly of compounds with two oxygenated positions in the steroidal A-ring (C-3, C-5). After feeding on P. ignitus males, Photuris females contain six major lucibufagins. Three of these compounds are not present in the prey, including the novel lucibufagin glycoside 5β,11α-dihydroxy-12-oxo-3β-O-β-D-xylopyranosylbufalin, and two other lucibufagins with a trioxygenated A-ring (C-2, C-3, C-5). These results indicate that Photuris females transform the sequestered lucibufagins both by glycosylation and oxidation, which could affect the systemic transportability of these compounds due to an increase in their polarity. Received 18 February 1999; accepted 19 April 1999.     

Behavioural evidence for the involvement of Dufour’s gland secretion in nestmate recognition in the social wasp Polistes dominulus (Hymenoptera: Vespidae)

The results of recent studies by our group have suggested that in social wasps of the genus Polistes the Dufour’s gland is involved in kin recognition. In fact, the same hydrocarbons occurring on the cuticle are found in the gland secretion, and in P. dominulus the composition of the glandular secretion is more similar in foundresses belonging to the same colonies than in heterocolonial foundresses. In this study, P. dominulus colonies were experimentally presented with previously treated dead conspecific females. These lures had been deprived of their epicuticular lipids and coated with epicuticular or Dufour’s gland secretion extracts from females hetero- or homocolonial with respect to the tested colonies. The behaviour of the colonies towards these lures indicates that, like the epicuticular lipids, the Dufour’s gland secretion is involved in nestmate recognition. Received: 14 August 1995/Accepted after revision: 5 November 1995     

Reef coral reproduction in the Abrolhos Reef Complex, Brazil: the endemic genus Mussismilia

The current contribution deals with the reproductive biology of a genus endemic to Brazil –Mussismilia Ortmann, 1890 – including all three species of the genus: M. braziliensis (Verrill, 1868), M. hartti (Verrill, 1868), and M. hispida (Verrill, 1902), which occur sympatrically in the studied area, the Abrolhos Reef Complex, Brazil. Sexuality patterns, modes of reproduction, synchrony and spawning periods are reported, and were determined by histological examination of material. All three species started to develop female and male gametes over different periods in the same breeding season. The three species are probably broadcast spawners, since no embryos or planulae were observed in any species at any given time of the year. Each reproductive cycle lasted approximately 11 months. Oogenesis and spermatogenesis started in different periods, with spermaries appearing in approximately the eighth month of ovary development and lasting about 3 months. Reproductive cycles were annual. Spawning probably occurred in consecutive months in each species. In M. braziliensis, spawning presumably happened between March and the middle of May in 1996 and 1997. Evidence suggested that spawning events of M. hispida took place between the end of April and mid-June. M. hartti may have spawned between September and November. The data presented here suggested that all studied species have at least one exclusive spawning period, asynchronically with the others. A possible exception may be the simultaneous (or close) spawnings of M. braziliensis and M. hispida in May. It is suggested that asynchrony in spawning periods among species may reduce the chance of hybridization, gamete waste and the competition for settlement surfaces. The occurrence of extended spawning periods for each species may also reduce the risks of reproductive failure, due to temporary adverse conditions. Received: 8 December 1998 / Accepted: 15 July 1999     

Triterpenoid and caffeic acid derivatives in the leaves of ragweed, <Emphasis Type="Italic">Ambrosia artemisiifolia</Emphasis> L. (Asterales: Asteraceae), as feeding stimulants of <Emphasis Type="Italic">Ophraella communa</Emphasis> LeSage (Coleoptera: Chrysomelidae)

Summary. The leaf beetle Ophraella communa infests almost exclusively Ambrosia artemisiifolia in the fields of Japan, even though it normally feeds on several Asteraceous plants. A filter paper bioassay showed that the feeding of O. communa is strongly stimulated by methanolic extracts of A. artemisiifolia. The feeding stimulants for O. communa have been isolated from methanolic extracts of A. artemisiifolia. -Amyrin acetate, -amyrin acetate, 5-caffeoylquinic acid (chlorogenic acid) and 3,5-dicaffeoylquinic acid from A. artemisiifolia have been identified as feeding stimulants for O. communa. Triterpenoid derivatives (-amyrin acetate or -amyrin acetate) and caffeic acid derivatives (3, 5-dicaffoylquinic acid or 5-caffeoylquinic acid) showed feeding stimulant activity when mixed together.     

Seasonal abundance and toxicity of the dinoflagellate Gambierdiscus spp. (Dinophyceae), the causative agent of ciguatera in Tahiti, French Polynesia

The benthic dinoflagellate Gambierdiscus spp. is the primary causative agent of ciguatera fish poisoning in French Polynesia. However, the initiation of ciguatera outbreaks and the factors that control ciguatoxin production in this dinoflagellate are still poorly understood. In this paper, we present some original data concerning the seasonal abundance and toxicity of natural populations of Gambierdiscus spp. based on a long-term survey in a ciguateric site of the island of Tahiti. From February 1993 to December 1997, Gambierdiscus spp. population densities were monitored weekly in the Atimaono barrier reef of Papara in relation to temperature and salinity. Densities peaked at 4992 cells g⁻¹ in October 1994 and constituted ≥1000 cells g⁻¹ on several occasions. A total of 58 blooms were recorded in the area, of which 65% occurred in 1995 and 1996 alone. Our data confirmed the high endemicity of Gambierdiscus spp. in the Papara area. Refined analysis of our raw data by a principal-component analysis of processes (PCAP) revealed a seasonality in cell densities from February 1993 to May 1995. During this period, Gambierdiscus spp. populations tended to reach maximum abundance at the beginning and end of the hot season. In contrast, salinity did not appear to be a determining factor in the seasonal abundance of this dinoflagellate. The noticeable increase in both peak densities and frequency of blooms further noticed in 1995 and 1996 was preceded by unusually high water temperatures in January to April 1994, concomitant with a severe coral-bleaching episode. During the course of the study, a total of 303 × 10⁶ cells obtained from 48 successive blooms was harvested. Toxicity screening revealed that toxin production was maximum from October 1994 through December 1996. No correlation was found between toxicity of these blooms and their biomass nor the seasonal pattern of temperatures. It is suggested that the toxicity of naturally-occurring blooms of Gambierdiscus spp. and, consequently, the severity of ciguatera incidents in a given area, is mainly dependent on the clonal nature of cells which coexist within local populations of this dinoflagellate. Received: 12 October 1998 / Accepted: 15 June 1999     

Genetic divergence and phylogenetic inferences in five species of Mugilidae (Pisces: Perciformes)

Allozyme electrophoresis was used to compare the genetic divergence of global populations of Mugil cephalus Linnaeus, 1758 and two congeneric [M.␣curema Cuvier & Valenciennes, 1836; M. gyrans (Jordan & Gilbert, 1885)] and two more distantly related [Liza ramada (Risso, 1826); Xenomugil thoburni (Jordan & Starks, 1896)] species on the basis of 18 enzyme loci. The amount of genetic divergence among the species examined is in agreement with their present systematic status, the differences being larger among non-congeneric species than among species of the same genus. Intraspecific genetic distances in M. cephalus (average Nei's D= 0.154), although higher than those reported for conspecific populations of fish, appear to be small when compared to the interspecific values among mugilid species (0.821 ≤ Nei's D ≤ 1.744). Phylogenetic trees obtained by genetic distance methods and discrete character parsimony analysis were of similar topology, except for the relationships within the genus Mugil and for the arrangement of M. cephalus populations. Received: 7 April 1997 / Accepted: 4 February 1998     

Water-soluble vitamins in natural plankton (copepods) during two consecutive spring blooms compared to vitamins in Artemia franciscana nauplii and metanauplii

Weight-specific contents of asorbic acid, thiamin, riboflavin, niacin, pantothenic acid, vitamin B₆, biotin, folate and vitamin B₁₂ were analysed in natural zooplankton collected from a sea water lagoon in a halibut fry production plant. The samples of natural zooplankton were collected from April/May to the end of June during two consecutive spring blooms. For comparison, the same vitamins were measured in nauplii of Artemia franciscana, just after hatching and after enrichment for 24 and 48 h with a commercial diet. The changes in vitamin content in zooplankton, which consisted mainly of Temora longicornis, during the two spring blooms were <2.5-fold for each specific vitamin. The large number of high quality halibut fry produced using the plankton as a dietary source suggests that the plankton contained sufficient amounts of all water-soluble vitamins. With the exception of thiamin and ascorbic acid, Artemia– both newly hatched and enriched for 24 and 48 h – contained higher levels of water-soluble vitamins than natural zooplankton. Enrichment feeding of Artemia for 48 h stabilised or increased the level of the water-soluble vitamins. Received: 8 March 1999 / Accepted: 21 February 2000     

Phylogeny of Opsanus spp. (Batrachoididae) inferred from multiple mitochondrial-DNA sequences

 Species of the toadfish genus Opsanus are increasingly used as experimental organisms in biomedicine and evolutionary/ecological physiology. Despite their importance, little is known of the phylogenetic relationships among Opsanus species. DNA sequence data for portions of the mitochondrial 12S rRNA, cytochrome oxidase I and cytochrome b genes were generated and analyzed from the four recognized Opsanus species and three outgroup taxa. Results of these analyses indicate an evolutionarily rapid divergence of three lineages within the genus. O. beta and O. pardus were resolved within the same lineage, and could not be distinguished with these mitochondrial sequence data. These data and paleoclimatic theory support a hypothesis of speciation based on the formation of allopatric populations during periods of elevated water temperature in the northern Caribbean. Received: 6 July 1999 / Accepted: 15 March 2000     

DMSP-lyase activity in five marine phytoplankton species: its potential importance in DMS production

Activity of DMSP-lyase, which cleaves dissolved DMSP (henceforth DMSP_d-lyase), was examined in five axenically cultured phytoplankton species, including both DMSP-producing and non-DMSP-producing species. High DMSP_d-lyase activity was found in two DMSP producers, Heterocapsa triquetra strain NIES-7 and Scrippsiella trochoidea strain NIES-369 (Dinophyceae). The DMS production rates at 100 nM DMSP_d were 0.5 fmol cell⁻¹ min⁻¹ for H. triquetra and 0.3 fmol cell⁻¹ min⁻¹ for S. trochoidea. In a non-DMSP producer, Heterosigma akashiwo strain NIES-6 (Raphidophyceae), the DMSP_d-lyase activity was not found. Two DMSP-producing Prymnesiophyceae species, Isochrysis galbana strain CCMP-1323 and Gephyrocapsa oceanica strain NIES-353, did not show any obvious activity either, in contrary to other authors' findings on Phaeocystis sp., another DMSP-producing Prymnesiophyceae species. The comparison of the DMSP_d-lyase activity of the two Dinophyceae species with bacterial DMSP consumption and DMS production activity in Tokyo Bay showed that the DMSP_d-lyase activity of H. triquetra and S. trochoidea could be an important mechanism for DMS production during their blooms. Received: 9 April 1999 / Accepted: 10 December 1999     

Identification of settling and early postlarval stages of mussels (Mytilus spp.) from the Pacific coast of North America, using prodissoconch morphology and genomic DNA

 Detailed inventories of the benthos and field studies of the settlement and recruitment processes of marine benthic invertebrates require accurate identification of newly settled larvae and early juvenile stages. We provide morphological criteria, visible under a good quality dissecting stereomicroscope, by which to discriminate between species of the settling larval and early postlarval stages (∼250 to 700 μm shell length) of mussels of the genus Mytilus on the west coast of Vancouver Island and Southern California. Compared to the bay mussel (M. trossulus), the sea mussel (M. californianus) has: (i) a shallower and flatter umbo, the latter corresponding to a significantly less pronounced prodissoconch I (PI) curvature and (ii) a greater PI length; as well as (iii) a wider separation between the provincular lateral teeth (PLT). The PLT distance is a new term denoting the separation between the midpoint of two reddish pigment spots of the provinculum (larval hinge apparatus) region of settling larvae and early postlarvae of Mytilus spp. from the East Pacific Coast. These spots mark the larger provincular lateral teeth, situated at either end of the provinculum. We confirmed the validity of morphological criteria by comparing PCR products of genomic DNA of provisionally identified postlarvae. Furthermore, measurements of PI lengths and PLT distance from well-preserved postlarvae of sea mussels (M. californianus) and of bay mussels (M. galloprovincialis) from Southern California indicate that the PI morphology and morphometry, and PLT distance criterion apply for that region as well. The criteria presented here can also apply to the advanced (competent) veliger stages, as the latter may settle (i.e. become the “settling” stage) upon encountering a suitable substrate. Our present and previously published work provide economical and effective identification methods that can be used to discriminate among early life history stages (∼250 μm to 5.0 mm shell length) of Mytilus spp. along the west coast of North America. Received: 10 November 1999 / Accepted: 6 September 2000     

Optical–digital system applied to the identification of five phytoplankton species

An optical–digital system was used to identify five phytoplankton species of the genus Ceratium from Todos Santos Bay off the California Peninsula, Mexico. The system recognized species with at least 90% accuracy, regardless of their size, rotation, or location within the field of view. An optical filter was constructed for each species for correlation with each of six test images. The test images were prepared from phytoplankton samples containing one or more of the five species. The correlations demostrate that each optical filter recognized its corresponding species. To evaluate the effectiveness of the correlation technique, we used images of 100 different samples. The species of the Ceratium genus analyzed are important indicators, since some are responsible for red-tide blooms in areas surrounding Todos Santos Bay, Baja California, Mexico. In the near future, based on the method proposed, we plan to implement an automated plankton-identification system to determine marine eco-biological properties. Received: 9 June 1997 / Accepted: 3 June 1998     

Chemical egg defense in Photuris firefly “femmes fatales”

Summary. Female Photuris fireflies contain defensive chemicals of two types. They sequester steroidal pyrones (lucibufagins) from male fireflies of the genus Photinus that they eat, and themselves produce the defensive betaine N-methylquinolinium 2-carboxylate. Chemical analyses of Photuris eggs showed that females that fed on Photinus males endow their eggs with both lucibufagin and the betaine, while those that did not feed on Photinus lay eggs that contain betaine, but virtually no lucibufagin. Photuris females collected in the field during the Photinus flight season laid eggs that invariably contained betaine, but lucibufagin only at times. Predation experiments showed that Photuris eggs are essentially unacceptable to larvae of a coccinellid beetle (Harmonia axyridis) and an ant (Leptothorax longispinosus), but moderately acceptable to an earwig (Forficula auricularia). When applied experimentally to palatable insect eggs, lucibufagin proved deterrent to these three predators, while the betaine proved deterrent to the ant and coccinellid larva only. Both types of defensive compound decreased egg predation in the field. By endowing their eggs with both exogenous and endogenous chemicals, Photuris females are essentially “maximizing their options”– when feeding on Photinus, their eggs are doubly protected, but they are not entirely defenseless when the females are unable to procure lucibufagin. Received 12 August 1999; accepted 27 August 1999     

Cytogenetic analysis of Oedalechilus labeo (Pisces: Mugilidae), with a report of NOR variability

 A cytogenetic analysis by several staining techniques was carried out on Oedalechilus labeo specimens from the west coast of Italy, in order to extend the karyological knowledge on Mediterranean mullets. The karyotype of the species is composed of 46 acrocentric and 2 subtelocentric chromosomes. Constitutive heterochromatin was found to be restricted to the centromeres. Fluorochrome staining revealed a uniform base composition along the chromosome arms. Nucleolus organizer regions (NORs) are located on the short arms of chromosome pair 9 and can be either homomorphic or quite heteromorphic in size. In some specimens chromomycin A₃-staining and fluorescence in situ hybridization with 18S rDNA revealed a third additional and inactive NOR on the short arms of a medium-sized chromosome. This is the first report of NOR variability in Mugilidae, a family generally characterized by a quite conservative karyotype. Heterochromatin composition and NOR location differentiate O. labeo from a cytotaxonomical point of view from the Liza species studied, Liza being considered the genus from which the genus Oedalechilus derived. Received: 10 April 1999 / Accepted: 21 October 1999     

Leaf surface compounds of the potato (<Emphasis Type="Italic">Solanum tuberosum</Emphasis>) and their influence on Colorado potato beetle (<Emphasis Type="Italic">Leptinotarsa decemlineata</Emphasis>) feeding

Summary.  The sampling behavior of the Colorado potato beetle Leptinotarsa decemlineata Say (CPB) involves examination of the surface of potato leaves. It has been suggested that leaf surface compounds (volatiles and cuticular waxes) may be involved in host-plant recognition, acceptance or discrimination. Here we report on the effect of leaf surface extracts of six Polish commercial potato varieties on CPB feeding. We tested the influence of potato leaf surface extracts on CPB adult and larval feeding, then separated the extracts with HPLC, and finally tested the effect of the HPLC-separated fractions on CPB feeding. The bioassays were performed using potato leaf discs deprived of their original surface compounds. Applied to test discs at concentrations ten times higher than natural (10 leaf area equivalent), the extracts deterred CPB adults and larvae from feeding. HPLC-separated fractions composed of alkanes, sesquiterpene hydrocarbons, wax esters, benzoic acid esters, fatty acid methyl, ethyl, isopropyl and phenylethyl esters, aldehydes, ketones, methyl ketones, fatty acids, primary alcohols, β-amyrin and sterols did not affect adult CPB feeding. Similarly, alkanes, sesquiterpene hydrocarbons, wax esters, methyl ketones, sesquiterpene alcohols and secondary alcohols had no effect on larval CPB feeding. The sterol fraction (cholesterol, β-sitosterol and stigmasterol) acted as a phagostimulant to CPB larvae. We isolated a fraction demonstrating a phagodeterrent effect on CPB adults and larvae. The qualitative composition of the deterrent fraction was quite similar in all potato extracts, but there were quantitative differences between the varieties. Much further work is needed to identify the compounds that can produce the deterrent effect.     

Impact of a burrowing deposit-feeder, Monoporeia affinis, on viable zooplankton resting eggs in the northern Baltic Sea

We investigated the effect of different densities of the burrowing deposit-feeding amphipod Monoporeia affinis on the potential for recruitment of zooplankton from benthic resting eggs. Hatching of resting eggs was induced in the laboratory on sliced and resuspended 1-cm depth-sections of sediment cores, collected at six stations in an archipelago area of the Gulf of Bothnia, Baltic Sea. The uppermost 5 cm of the sediment was studied. The most common species that hatched was Eurytemora affinis (Copepoda). Individuals from another copepod genus, Acartia, hatched in significant numbers only in the cores from two stations with low amphipod abundance. Cores from stations with high amphipod densities showed a deeper distribution of emerging E. affinis nauplii compared with stations with few amphipods; the oxidised sediment layer was also deeper at high M. affinis densities than at low. Total (0 to 5 cm strata pooled) number of hatched E. affinis nauplii was independent of amphipod density. This indicates that the effect of M. affinis on E. affinis eggs involves deeper burial due to bioturbation, rather than predation. Decreased benthic recruitment of zooplankton at localities with high M. affinis density is suggested, since more deeply positioned eggs are less likely to hatch. When hatching was induced in intact, non-sliced cores from one station, the number of E. affinis nauplii that hatched was on average 43% of the number that hatched in the upper centimetre of the sliced cores from the same station. This fraction (43%), if applied to the other stations, implied a potential for benthic recruitment of up to 80 000 ind m⁻² for E. affinis. Due to its high abundance, M. affinis is likely to greatly reduce benthic recruitment of zooplankton in this system. Received: 17 September 1999 / Accepted: 23 February 2000     

Effect of host and larval frass volatiles on behavioural response of the old house borer,Hylotrupes bajulus (L.) (Coleoptera: Cerambycidae), in a wind tunnel bioassay

Summary. In a wind tunnel bioassay the effect of three concentrations of natural extracts of (1) Scots pine wood, Pinus sylvestris, and (2) larval frass on the behavioural response of unmated females and males of the old house borer, Hylotrupes bajulus, was tested and compared to the behavioural effects of the male-produced sex pheromone (3R)-3-hydroxy-2-hexanone. The influence on the behaviour of both sexes was found to be equally significant for the two higher concentrated hexane extracts of wood and larval frass. Therefore several synthetic monoterpenes present in the extracts and ethanol were tested at the two higher concentrations (1:100, 1:1000 vol/vol). Among the higher concentrated monoterpenoid hydrocarbons [(+)-α-pinene, (+)-β-pinene, (+)-limonene], only α-pinene increased the activity, orientation towards scent source and interest towards conspecifics. The tests with higher concentrated ethanol and the oxygenated monoterpenes [(-)-verbenone, (-)-trans-pinocarveol, (+)-terpinen-4-ol, (+)-α-terpineol, (-)-myrtenol] revealed that verbenone is the most effective stimulant for the females, followed by trans-pinocarveol, terpinen-4-ol and α-terpineol. For males, terpinen-4-ol was the only mediator significantly inducing attraction and orientation towards the scent source combined with an interest in conspecifics apparent by fighting or courtship behaviour. Males did not respond to verbenone which is a main compound of larval frass. Myrtenol and ethanol were ineffective in both sexes. In fact behavioural observations suggest that the beetles were repelled by the high dose of myrtenol. Using the ten-fold lower dose of the synthetic monoterpenes (1:1000 vol/vol), all semiochemicals except myrtenol lost activity. Myrtenol, however, induced behavioural responses, like increased activity and orientation towards scent source, only at the low concentration. Based on the results, primary attraction of unmated old house borer is probably mediated by monoterpenes of coniferous wood, while secondary attraction to infested wood would occur in response to volatiles of larval frass. Received 5 May 1999; accepted 30 September 1999     

<Emphasis Type="Italic">Cochlodinium polykrikoides</Emphasis> blooms and clonal isolates from the northwest Atlantic coast cause rapid mortality in larvae of multiple bivalve species

Globally, many commercial bivalve populations have declined in recent decades. In addition to overharvesting and habitat loss, the increasing frequency and intensity of harmful algal blooms (HABs) are likely to contribute to bivalve losses, particularly in cases where blooms negatively impact larval stages. This paper reports on the lethal effects of clonal cultures and blooms of Cochlodinium polykrikoides from the US Atlantic coast on the larvae of three species of commercially and ecologically valuable bivalves: the Eastern oyster (Crassostrea virginica), the bay scallop (Argopecten irradians), and the Northern quahog (hard clam; Mercenaria mercenaria). Both cultures and blooms of C. polykrikoides were highly toxic to all three species of bivalve larvae causing 80–100% mortality during 24- to 72-h exposures at concentrations of 1–2 × 10³ cells ml⁻¹. Toxicity was dependent on cell densities, growth stage of C. polykrikoides (i.e. cultures in exponential stage growth were more toxic than later stages), exposure time of larvae to cells (i.e. longer exposure caused higher mortality), the age of larvae (i.e. younger larvae were more sensitive), and the relative abundance of C. polykrikoides (i.e. the presence of other microalgae decreased toxicity). Free radical-scavenging enzymes (peroxidase and catalase) and the removal of C. polykrikoides cells (i.e. culture filtrate) significantly increased larval survival suggesting toxicity is maximized by contact with live cells and may involve labile toxins bound by these compounds including e.g. reactive oxygen species. The toxicity of C. polykrikoides to bivalve larvae was generally more severe than other HAB species (e.g. Karenia brevis, Karlodinium veneficum, Alexandrium tamarense, Prorocentrum minimum). Since the bivalves in this study spawn in the months when C. polykrikoides blooms on the east coast of North America, these results suggest that these blooms may have detrimental effects on efforts to restore these already diminished populations.     

Importance of cysts in the population dynamics of the red tide flagellate Heterosigma akashiwo (Raphidophyceae)

To elucidate roles of cysts in occurrences of Heterosigma akashiwo blooms, cyst dynamics were studied in northern Hiroshima Bay, the Seto Inland Sea of Japan, where H. akashiwo regularly forms red tide in June. Monthly measurements of seasonal changes in the densities of vegetative cells of H. akashiwo and their germinable cysts in surface sediments (top 1-cm layer) were made for 2 years at three stations. Vegetative cells of H. akashiwo could be detected from April through December throughout the water column, and the existence of vegetative cells was confirmed in surface waters even in winter after incubation of sampled seawater in culture medium. Germinable cysts, enumerated by the extinction dilution method, existed in sediments in all seasons, even before and after the seasonal bloom. The effects of incubation temperature on the germination of natural cysts of H. akashiwo in sediments were examined. Germination was not observed at 5 °C, was low at 10 °C, while it increased at 15 °C, and maintained a high level to 25 °C. The bottom water temperature reached 15 °C (suitable for the germination of cysts) and the surface about 18 °C or more (suitable for the growth of vegetative cells) 2 to 3 weeks before the blooms. The dark survival of H. akashiwo cysts was tested, and it was found that the cysts were viable for at least 650 d at 11 °C, and for 165 d at 25 °C, indicating a significant role of cysts in the survival during winter and summer seasons. The cysts presumably also play an important role in seeding primary populations into water columns when the bottom water reaches a suitable temperature (around 15 °C); thereafter the populations develop with great annual regularity to bloom in June. These results suggest that initiation of H. akashiwo red tides in the Seto Inland Sea could be triggered by bottom water temperature. Received: 3 July 1998 / Accepted: 12 January 1999     

Quantitative analysis of pyrrolizidine alkaloids sequestered from diverse host plants in Longitarsus flea beetles (Coleoptera, Chrysomelidae)

Summary. Several species of the flea beetles genus Longitarsus are able to sequester pyrrolizidine alkaloids (PAs) from their host plants. In five Longitarsus species we compare the concentration of PAs present in their host plants belonging to the Asteraceae or Boraginaceae with those found in the beetles. To get an estimate of the intrapopulation variability, three samples of five beetles each and five individual plants were analyzed for each comparison. A strong intrapopulation variability could be detected both among plant and beetle samples. The total concentration found in the beetles varied strongly between species. The local host plant and its phenology influence the concentrations present in the beetles as evidenced in comparisons of a single beetle species from two different hosts and of one beetle species collected at the same site at different times of the year. In addition, different beetle species apparently vary in their capacity to sequester the alkaloids, at the lowest extreme the mean PA concentration in the beetles (0.034 μg PA/mg dry weight) was 1/30 of the mean concentration found in the plant leaves (L. aeruginosus from Eupatorium cannabinum), at the highest extreme (2.098 μg PA/mg dw) the concentration in the beetles was a 1000 fold higher than in the plant leaves (L. nasturtii from Symphytum officinale). The highest mean concentration found in the beetles was 3.446 μg/mg dw (L. exoletus from Cynoglossum officinale). The absolute concentrations found in the beetles are comparable to other insects which have been shown to be effectively defended against their potential predators. Received 22 June 1999; accepted 25 August 1999     

Phytoplankton blooms are strongly impacted by microzooplankton grazing in coastal North Pacific waters

Phytoplankton growth and microzooplankton grazing were measured in two productive coastal regions of the North Pacific: northern Puget Sound and the coastal Gulf of Alaska. Rates of phytoplankton growth (range: 0.09–2.69 day⁻¹) and microzooplankton grazing (range: 0.00–2.10 day⁻¹) varied seasonally, with lowest values in late fall and winter, and highest values in spring and summer. Chlorophyll concentrations also varied widely (0.19–13.65 μg l⁻¹). Large (>8 μm) phytoplankton cells consistently dominated phytoplankton communities under bloom conditions, contributing on average 65% of total chlorophyll biomass when chlorophyll exceeded 2 μg l⁻¹. Microzooplankton grazing was an important loss process affecting phytoplankton, with grazing rates equivalent to nearly two-thirds (64%) of growth rates on average. Both small and large phytoplankton cells were consumed, with the ratio of grazing to growth (g:μ) for the two size classes averaging 0.80 and 0.42, respectively. Perhaps surprisingly, the coupling between microzooplankton grazing and phytoplankton growth was tighter during phytoplankton blooms than during low biomass periods, with g:μ averaging 0.78 during blooms and 0.49 at other times. This tight coupling may be a result of the high potential growth and ingestion rates of protist grazers, some of which feed on bloom-forming diatoms and other large phytoplankton. Large ciliates and Gyrodinium-like dinoflagellates contributed substantially to microzooplankton biomass at diatom bloom stations in the Gulf of Alaska, and microzooplankton biomass overall was strongly correlated with >8 μm chlorophyll concentrations. Because grazing tended to be proportionally greater when phytoplankton biomass was high, the absolute amount of chlorophyll consumed by microzooplankton was often substantial. In nearly two-thirds of the experiments (14/23), more chlorophyll was ingested by microzooplankton than was available for all other biological and physical loss processes combined. Microzooplankton were important intermediaries in the transfer of primary production to higher trophic levels in these coastal marine food webs. Received: 12 November 1999 / Accepted: 4 October 2000